3-quinolinecarbonitrile protein kinase inhibitors

ABSTRACT

This invention provides a compound of Formula 1 
     
       
         
         
             
             
         
       
     
     where Ar, X, R 1 , R 2 , R 3 , and R 4  are defined herein, or a pharmaceutically acceptable salt thereof useful in the prevention or inhibition of diseases associated with the Ras/Raf/MEK signaling cascade in a mammal, such as neoplasms, strokes, osteoporosis, cancer, rheumatoid arthritis, inflammatory disease, polycystic kidney disease, and colonic polyps, and methods of making the compounds of formula 1 and intermediates.

This application claims priority from copending provisional applicationSer. No. 60/546,511, filed Feb. 20, 2004, the entire disclosure of whichis hereby incorporated by reference.

BACKGROUND OF THE INVENTION

This invention relates to substituted 3-quinolinecarbonitrile compoundsas well as the pharmaceutically acceptable salts thereof. The compoundsof the present invention inhibit the action of certain enzymes known asprotein kinases. Protein kinases play a vital role as key regulators ofa variety of critical cell functions. These enzymes function bycatalyzing the transfer of a phosphate group from ATP to amino acidresidues of substrate proteins. Tumorigenesis has been linked to theaberrant function of protein kinases and protein kinases are ofparticular interest as potential targets for anticancer agents.

Protein kinases are a class of enzymes that catalyze the transfer of aphosphate group from ATP to a tyrosine, serine, threonine, or histidineresidue located on a protein substrate. Protein kinases clearly play arole in normal cell growth. Many of the growth factor receptor proteinsfunction as kinases and it is by this process that they effectsignaling. The interaction of growth factors with these receptors is anecessary event in normal regulation of cell growth. However, undercertain conditions, as a result of either mutation or over expression,these receptors can become deregulated. Specific protein kinases havebeen implicated in diverse conditions including cancer (Traxler, P. M.,Exp. Opin. Ther. Patents, 8, 1599 (1998); Bridges, A. J., EmergingDrugs, 3, 279 (1998)), restenosis (Mattsson, E., Trends Cardiovas. Med.5, 200 (1995); Shaw, Trends Pharmacol. Sci. 16, 401 (1995)),atherosclerosis (Raines, E. W., Bioessays, 18, 271 (1996)), angiogenesis(Shawver, L. K., Drug Discovery Today, 2, 50 (1997); Folkman, J., NatureMedicine, 1, 27 (1995)) and osteoporosis (Boyce, J. Clin. Invest., 90,1622 (1992)).

One signaling pathway found in all eukaryotic organisms is the Ras-MAPKmodule, which is comprised of the Ras/Raf/MEK (mitogen-activated proteinkinase kinase)/MAPK (mitogen-activated protein kinase) signalingcascade. This key pathway is involved in transmitting signals fromgrowth factors and hormones at the extracellular compartment into thecytosol and to transcription factors in the nucleus. Alteration of theRas-MAPK pathway is associated with the formation of certain humantumors.

Components of the Ras-MAPK Signaling Cascade

The Ras/Raf/MEK/MAPK signaling cascade is activated by GTP loading ofRas, which occurs in response to stimuli from cell surface receptors(Malumbres, M., Barbacid, M. RAS oncogenes: the first 30 years. Nat RevCancer 2003, 3: 459-65; Downward, J. Targeting RAS signalling pathwaysin cancer therapy. Nat Rev Cancer 2003, 3: 11-22; Bos, J. L. Rasoncogenes in human cancer. A review. Cancer Res 1989, 49: 4682-4689).Ras is a member of a large family of small (21 kDa) GTPases that act asmolecular switches in the regulation of cell growth, differentiation,survival, and apoptosis (Malumbres, M., Barbacid, M. RAS oncogenes: thefirst 30 years. Nat Rev Cancer 2003, 3: 459-65; Downward, J. TargetingRAS signalling pathways in cancer therapy. Nat Rev Cancer 2003, 3:11-22; Bos, J. L. Ras oncogenes in human cancer: A review. Cancer Res1989, 49: 4682-4689; Shields, J. M., Pruitt, K., McFall, A. et al.Understanding Ras: ‘it ain't over til it's over’. Trends Cell Biol 2000,10: 147-154). Ras is initially defined as a component of oncogenicmurine retroviruses (Ellis, R. W., Defeo, D., Shih, T. Y. et al. The p21src genes of Harvey and Kirsten sarcoma viruses originate from divergentmembers of a family of normal vertebrate genes. Nature 1981, 292:506-511), and it became the focus of intensive research in the early1980s when the connection between mutant forms of Ras and human cancerwas established (Ellis, R. W., Defeo, D., Shih, T. Y. et al. The p21 srcgenes of Harvey and Kirsten sarcoma viruses originate from divergentmembers of a family of normal vertebrate genes. Nature 1981, 292:506-511; Perucho, M., Goldfarb, M., Shimizu, K., Lama, C., Fogh, J. andWigler, M. Human-tumor-derived cell lines contain common and differenttransforming genes. Cell 1981, 27: 467-476; Santos, E., Martin-Zanca,D., Reddy, E. P., Pierotti, M. A., Della Porta, G., Barbacid, M.Malignant activation of a K-ras oncogene in lung carcinoma but not innormal tissue of the same patient. Science 1984, 223: 661-664). Rasmutations that lead to oncogenic activation occur primarily in twohotspots: Gly12 and Gln61 (Krengel, U., Schlichting, L., Scherer, A. etal. Three-dimensional structures of H-ras p21 mutants: Molecular basisfor their inability to function as signal switch molecules. Cell 1990,62: 539-548). The mutational changes at these residues found in humancancers impede the GTP hydrolysis activity of Ras, thus causing Ras toremain in the GTP-bound or “on” conformation. With this finding, abetter appreciation of the molecular basis of aberrant signalingassociated with ˜30% of all human cancers are obtained.

There are three Ras isoforms associated with human cancer, H-Ras, N-Ras,and K-Ras; and of these, 95% are due to K-Ras mutations. Ras proteinsare closely related, having 85% amino acid identity with a 20 amino acidvariable region at the carboxy terminus (Lowy, D. R. and Willumsen, B.M. Function and regulation of ras. Annual Review of Biochemistry 1993,62: 851-891). A Cysteine residue occurs in all Ras proteins after thevariable region, and this is the site of post-translational modificationof Ras by addition of a farnesyl isoprenoid lipid (Hancock, J. F.,Magee, A. I., Childs, J. E. and Marshall, C. J. All ras proteins arepolyisoprenylated but only some are palmitoylated. Cell 1989, 57:1167-1177). All Ras proteins are further modified by proteolysis of thethree carboxy terminal amino acids and subsequent methylation of the newcarboxy terminus. These modifications stabilize Ras interaction with theinner cell membrane where it must reside to form a multi-protein complexwith Raf, MEK, MAPK and scaffold proteins to activate signaling in theRas-MAPK module (Kolch, W. Meaningful relationships: The regulation ofthe Ras/Raf/MEK/ERK pathway by protein interactions. Biochemical J 2000,351 Pt 2: 289-305; Kolch, W. Ras/Raf signalling and emergingpharmacotherapeutic targets. Expert Opin Pharmaco 2002, 3: 709-718).

As the consequences of mutated Ras function were elucidated, thisattracted sufficient attention from numerous pharmaceutical laboratoriesto initiate discovery and development programs for small moleculeinhibitors of this signaling protein. The goal of these programs is tobring forward inhibitors of aberrant Ras signaling that have minimalassociated toxicities. Numerous Ras inhibitors have been described, andthey impair Ras-MARK signaling by blocking proper post-translationalmodification of Ras by protein farnesyl transferase, thereby preventingmembrane localization. These Ras inhibitors have shown good pre-clinicalefficacy, and are currently being evaluated in clinical trials. Thesequential downstream kinase effectors of Ras, namely Raft MEK, and MAPKare in addition viewed as equally attractive targets for thepharmacological intervention of cancer.

Rat proteins are fairly homologous (˜60%). These 66-84 kDaserine/threonine kinases include A, B, and C isoforms (C-Raf=Raf1,c-Raf), which coexist in many cell types and activate MEK (Kolch, W.Ras/Raf signalling and emerging pharmacotherapeutic targets. Expert OpinPharmaco 2002, 3: 709-718; Chong, H., Vikis, H. G., Guan, K.-L.Mechanisms of regulating the Raf kinase family. Cell Signal 2003, 15:463-469; Chong, H., Lee, J., Guan, K. L. Positive and negativeregulation of raf kinase activity and function by phosphorylation. EMBOJ 2001, 20: 3716-3727). Three conserved regions occur in Raf proteins:CR1 which is at the N-terminal and contains a Ras binding domain (RBD)and a cysteine rich domain (CRD); CR2 which contains a serine/threoninerich region; and CR3 which contains the catalytic kinase domain.GTP-loaded Ras recruits Raf to the inner cell membrane. This is crucialfor Raf activation, though activation is a complex process not yet fullyunderstood. For example, there are at least 13 regulatoryphosphorylation sites on C-Raf (Fabian, J. R., Daar, I. O., Morrison, D.K. Critical tyrosine residues regulate the enzymatic and biologicalactivity of raf-1 kinase. Mol Cell Biol 1993, 13: 7170-7179; Marais, R.,Wynne, J., Treisman, R. The SRF accessory protein Elk-1 contains agrowth factor-regulated transcriptional activation domain. Cell 1993,73: 381-393; Morrison, D. K., Heidecker, G., Rapp, U. R. and Copeland,T. D. Identification of the major phosphorylation sites of the Raf-1kinase. J Biol Chem 1993, 268: 17309-17316; Mason, C. S., Springer, C.J., Cooper, R. G., Superti-Furga, G., Marshall, C. J., Marais, R. Serineand tyrosine phosphorylations cooperate in Raf-1 but not B-Rafactivation. EMBO J 1999, 18: 2137-2148; Dhillon, A. S., Meikle, S.,Yazici, Z., Eulitz, M., Kolch, W. Regulation of Raf-1 activation andsignalling by dephosphorylation. EMBO J 2002, 21: 64-71; Kolch, W. To beor not to be: a question of B-Raf? Trends Neurosci 2001, 24: 498-600;Abraham, D., Podar, K., Pacher, M. et al. Raf-1-associated proteinphosphatase 2A as a positive regulator of kinase activation. J Biol Chem2000, 275: 22300-22304; Yeung, K., Seitz, T., Li, S. et al. Suppressionof Raf-1 kinase activity and MAP kinase signalling by RKIP. Nature 1999,401: 173-177; Chang, F., Steelman, L. S., Lee, J. T. et al. Signaltransduction mediated by the RasRafMEKERK pathway from cytokinereceptors to transcription factors: potential targeting for therapeuticintervention. Leukemia 2003 17:1263-1293). Not only GTP-loaded Ras, butalso various kinases (e.g. C-Tak1, PAK, PKC, PKA, Src), phosphatases(PP1, PP2A), adapter proteins, and scaffold proteins (KSR) areimplicated in full Raf activation (Dhillon, A. S. and Kolch, W. Untyingthe regulation of the Raf-1 kinase. Arch Biochem Biophys 2002, 404: 3-9;Morrison, D. K. KSR: a MAPK scaffold of the Ras pathway? J Cell Sci2001, 114: 1609-1612; Baccarini, M. An old kinase on a new path: Raf andapoptosis. Cell Death Differ 2002, 9: 783-785). The three Raf isoformsdiffer in their ability to interact with Ras isoforms, to activate MEK,and to transform rodent fibroblasts in vitro (Pritchard, C. A., Bolin,L., Slattery, R., Murray, R., McMahon, M. Post-natal lethality andneurological and gastrointestinal defects in mice with targeteddisruption of the A-Raf protein kinase gene. Curr Biol 1996, 6:614-617). The B-Raf isoform in all cases is the most active followed byC-Raf, and then A-Raf.

A, B, and C Raf knockout mice have been described (Wojnowski, L.,Zimmer, A. M., Beck, T. W. et al. Endothelial apoptosis inBraf-deficient mice. Nat Genet 1997, 16: 293-297; Huser, M., Luckett,J., Chiloeches, A. et al. MEK kinase activity is not necessary for Raf-1function. EMBO J 2001, 20: 1940-1951; Murakami, M. S., Morrison, D. K.Raf-1 without MEK? Science's Stke: Signal Transduction KnowledgeEnvironment 2001, 2001: PE3). B-Raf deficient embryos die atmid-gestation due to apoptotic cell death in endothelial cells leadingto vascular hemorrhage. C-Raf deficiency causes mid-gestational deathdue to more diffuse apoptotic tissue effects. A-Raf deficient mice areborn alive, but show neurological and intestinal defects. Thesedivergent phenotypes show that Raf isoforms serve distinct functions indifferent tissues. These studies have shown that individual B-Raf andC-Raf survival functions cannot be performed by other Raf isoforms. Theyalso demonstrated that normal levels of MAPK activation occurs in C-Rafdeficient mouse cells, indicating that the anti-apoptotic function ofC-Raf is not mediated by the MAPK cascade.

The anti-apoptotic function of C-Raf may be mediated by antagonism ofapoptosis-stimulated kinase 1 (ASK-1). There is evidence to suggest thatC-Raf impedes ASK-1 function via a protein-protein interaction that isnot associated with C-Raf kinase activity (Chen, J., Fujii, K., Zhang,L., Roberts, T., Fu, H. Raf-1 promotes cell survival by antagonizingapoptosis signal-regulating kinase 1 through a MEK-ERK independentmechanism. Proc Natl Acad Sci USA 2001, 98: 7783-7788). Raf also canimpede apoptosis in a kinase dependent manner. For example, Raf/MEK/MAPKsignaling activates Rsk1, which in turn phosphorylates and inactivatesBAD, a pro-apoptotic protein (Shimamura, A., Ballif, B. A., Richards, S.A., Blenis, J. Rsk1 mediates a MEK-MAP kinase cell survival signal. CurrBiol 2000, 10: 127-135). Additionally, C-Raf can be localized to themitochondria by a Bcl-2 mediated process, where it can inactivatepro-apoptotic proteins by phosphorylation (Wang, H. G., Miyashita, T.,Takayama, S. et al. Apoptosis regulation by interaction of Bcl-2 proteinand Raf-1 kinase. Oncogene 1994, 9: 2751-2756). Raf anti-apoptoticeffects are complex and will require further study to clarify. However,this characteristic of Raf enhances its appeal as a pharmaceuticaltarget since a hallmark of cancer cells is resistance to apoptosis whichat least in part is likely attributable to improper Raf activation(Herrera, R., Sebolt-Leopold, J. S. Unraveling the complexities of theRaf/MAP kinase pathway for pharmacological intervention. Trends Mol Med2002, 8: S27-31).

As with Ras, oncogenic forms of Raf have been found to be components oftransforming murine retroviruses (Mark, G. E., Rapp, U. R. Primarystructure of v-raf: relatedness to the src family of oncogenes. Science1984, 224: 285-289). Oncogenic Raf in murine retroviruses results fromN-terminal deletions that remove the regulatory sequences that controlRaf kinase activity. Most recently, a systematic human genome-widescreening effort to detect alterations in genes that control cellproliferation, differentiation, and death found activating B-Rafmutations in 66% of malignant melanomas (Davies, H., Bignell, G. R.,Cox, C. et al. Mutations of the BRAF gene in human cancer. Nature 2002,417: 949-954). Additionally, B-Raf mutations are observed at lowerfrequencies in a wide range of other cancers including colorectal, lung,breast and ovarian.

The significance of B-Raf mutations in colorectal tumors is extended ina subsequent study showing that mutations in either B-Raf or K-Ras (notboth) are detected in a sample of colorectal tumors examined at allstages of development, including pre-malignant lesions (Rajagopalan, H.,Bardelli, A., Lengauer, C., Kinzier, K. W., Vogetstein, B., Velculescu,V. E. Tumorigenesis: RAF/RAS oncogenes and mismatch-repair status.Nature 2002, 418: 934). The frequency of B-Raf mutations in thesecolorectal tumors is 10%, whereas the K-Ras mutation frequency is 51%.Cumulative K-Ras/B-Raf mutation frequency in colorectal cancer istherefore 61%. In the case of melanoma, an examination of N-Ras mutation(the Ras isoform mutated in melanoma), together with B-Raf has shown acumulative mutation frequency of 81% (Smalley, K. S. A pivotal role forERK in the oncogenic behaviour of malignant melanoma? Int J Cancer 2003,104: 527-532). These statistics, when combined with the 2002 incidenceof colorectal cancer (148,000) and melanoma (54,000) in the U.S.(American Cancer Society), make Raf a compelling pharmaceutical target.

MEK1 and MEK2 are expressed 43-46 kDa kinases activated by Rafphosphorylation of two serine residues (Ser217-Ser221). MEK1 and MEK2are members of a larger family of dual specificity kinases (MEK1-7) thatphosphorylate Threonine and Tyrosine residues within the TXY motif ofvarious MAP kinases (Dhanasekaran, N., Premkumar Reddy, E. Signaling bydual specificity kinases. Oncogene 1998, 17: 1447-1455). MEK1 and MEK2are encoded by distinct genes, but they have high homology (80%) withinthe C-terminal catalytic kinase domain and most of the N-terminalregulatory domain (English, J., Pearson, G., Wilsbacher, J. et al. Newinsights into the control of MAP kinase pathways. Exp Cell Res 1999,253: 255-270). At the N-terminus of MEK1 and MEK2 there are 30 aminoacids of divergent sequence that may direct differential interactionswith both activators and substrates. The only known substrates forMEK1/MEK2 are the MAPK1 and MAPK2, which they phosphorylate onThr202/183 and Tyr204/185, respectively.

MEK1 deficient mice have been described, and inactivation of MEK1 leadsto embryonic lethality due to decreased placental vascularization duringembryogenesis (Giroux, S., Tremblay, M., Bernard, D. et al., Embryonicdeath of Mek1-deficient mice reveals a role for this kinase inangiogenesis in the labyrinthine region of the placenta. Curr Biol 1999,9: 369-372). MEK1 deficiency is not compensated for by MEK2. Incontrast, MEK2 deficient mice are viable and fertile, with nomorphological alterations (Bélanger, L. F., Roy, S., Tremblay, M. et al.Mek2 is dispensable for mouse growth and development Mol Cell Biol 2003,23: 4778-4787). These data demonstrate that MEK2 is not necessary forthe normal development of mouse embryos, indicating that the loss ofMEK2 can be compensated for (Bélanger, L. F., Roy, S., Tremblay, M. etal. Mek2 is dispensable for mouse growth and development. Mol Cell Biol2003, 23: 4778-4787) by at least in part MEK1.

Oncogenic forms of MEK1 or MEK2 have not been described in retrovirusesor human cancers. However, a MEK1 where Ser218 and Ser222 are bothmutated to Asp is capable of causing oncogenic transformation of variousrodent fibroblast cell lines (Mansour, S. J., Matten, W. T., Hermann, A.S. et al. Transformation of mammalian cells by constitutively active MAPkinase. Science 1994, 265: 966-970).

The MAPK components of the Ras-MAPK module have also been designatedERK1 and ERK2 (extracellular signal-regulated kinases). These MAPKisoforms (also designated p44 MAPK and p42 MAPK) are highly homologous(>80%), expressed 44-42 kDa serine/threonine kinases that are members ofa larger gene family that includes ERK 1, 2, 3, 5, 7; JNK 1-3; and p38

and

. Experimental data indicate that ERK1 and ERK2 are functionallyequivalent (English, J. M., Cobb, M. H. Pharmacological inhibitors ofMAPK pathways. Trends Pharmacol Sci 2002, 23: 40-45). ERKs are activatedby MEK phosphorylation of their TEY sequence; dual phosphorylation isrequired for activation, and in the case of ERK2 results in a >1000 foldincrease in activity. Downstream substrates of ERK1/2 includecytoskeletal proteins, kinases, phosphatases, and transcription factors.The pleiotropic effects of MAPK activation on cell growth anddifferentiation are undoubtedly mediated through this diverse array ofeffectors.

No constitutively active MAP kinases are known, despite attempts attheir genetic selection and site-directed mutagenesis. This failuresuggests that cells cannot tolerate the continuous activation of MAPkinases. Among the kinase components of the Ras-MAPK signaling only theERK2 atomic structure has been solved (Zhang, F., Strand, A., Robbins,D., Cobb, M. H., Goldsmith, E. J. Atomic structure of the MAP kinaseERK2 at 2.3 Á resolution. Nature 1994, 367: 704-711).

The ERK1/ERK2 components of the Ras-MAPK module are the most abundant(˜10⁶ molecules per cell). MEK also is relatively abundant in most celltypes (˜3.5×10⁵ molecules per cell), whereas Raf and Ras molecules areless abundant (˜2×10⁴ per cell) (Ferrell, J. E., Jr. Tripping the switchfantastic: how a protein kinase cascade can convert graded inputs intoswitch-like outputs. Trends Biochem Sci 1996, 21: 460-466). All MAPKmolecules can become fully activated in cells where only 10-50% of Rasmolecules are GTP bound. The predicted sensitivity of the Raf/MEK1/MAPKsignaling cascade to inhibitors is: Raf>MEK1>MAPK (Huang, C. Y.,Ferrell, J. E., Jr. Ultrasensitivity in the mitogen-activated proteinkinase cascade. Proc Natl Acad Sci USA 1996, 93: 10078-10083). Thissensitivity profile results from the distributive (non-processive)mechanism of both Raf and MEK1 in which the rate of MEK1 activationdepends on the concentration of Raf squared; and similarly the rate ofMAPK activation is dependent on the concentration of MEK1 squared. Todate, potent inhibitors of Raf and MEK, but not ERK, have been reported.

BRIEF SUMMARY OF THE INVENTION

This invention relates to substituted 3-quinolinecarbonitrile compoundsuseful for the treatment or inhibition of certain diseases that are theresult of deregulation of these protein kinases. The compounds of thisinvention are anti-cancer agents and are useful for the treatment orinhibition of cancer in mammals. In addition, the compounds of thisinvention are useful for the treatment and inhibition of stroke,osteoporosis, rheumatoid arthritis and other inflammatory disorders, aswell as polycystic kidney disease and colonic polyps.

In accordance with this invention there is provided a group of compoundsrepresented by Formula 1:

wherein:Ar is a cycloalkyl of 3 to 7 carbon atoms, which may be optionallysubstituted with one or more alkyl of 1 to 6 carbon atoms, a pyridinyl,a pyrimidinyl, or a phenyl ring; wherein the pyridinyl, pyrimidinyl, orphenyl ring may be optionally mono-, di-, or tri-substituted withsubstituents selected from a group consisting of a halogen, an alkyl of1-6 carbon atoms, an alkenyl of 2-6 carbon atoms, an alkynyl of 2-6carbon atoms, azido, a hydroxyalkyl of 1-6 carbon atoms, a halomethyl,an alkoxymethyl of 2-7 carbon atoms, an alkanoyloxymethyl of 2-7 carbonatoms, an alkoxy of 1-6 carbon atoms, an alkylthio of 1-6 carbon atoms,a hydroxy, a trifluoromethyl, a cyano, a nitro, a carboxy, analkoxycarbonyl of 2-7 carbon atoms, an alkanoyl of 2-7 carbon atoms, abenzoyl, an amino, an alkylamino of 1-6 carbon atoms, a dialkylamino of2 to 12 carbon atoms, an alkanoylamino of 1-6 carbon atoms, analkenoylamino of 3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms,an alkanoyloxy of 1-6 carbon atoms, an alkenoyloxy of 3-8 carbon atoms,an alkynoyloxy of 3-8 carbon atoms, a carbamoyl, an N-alkylcarbamoyl of2-7 carbon atoms, a N,N-dialkylcarbamoyl of 3-13 carbon atoms, acarboxyalkyl of 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbonatoms, an aminoalkyl of 1-5 carbon atoms, an N-alkylaminoalkyl of 2-9carbon atoms, a N,N-dialkylaminoalkyl of 3-10 carbon atoms, aN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, mercapto, methylmercapto and benzoylamino;a bicyclic aryl or bicyclic heteroaryl ring system of 8 to 12 atomswhere the bicyclic heteroaryl ring contains 1 to 4 heteroatoms selectedfrom N, O, and S, wherein the bicyclic aryl or bicyclic heteroaryl ringmay be optionally mono- di-, tri, or tetra-substituted with asubstituent comprising a halogen, an oxo, a thiocarbonyl, an alkyl of1-6 carbon atoms, an alkenyl of 2-6 carbon atoms, an alkynyl of 2-6carbon atoms, azido, a hydroxyalkyl of 1-6 carbon atoms, a halomethyl,an alkoxymethyl of 2-7 carbon atoms, an alkanoyloxymethyl of 2-7 carbonatoms, an alkoxy of 1-6 carbon atoms, an alkylthio of 1-6 carbon atoms,a hydroxy, a trifluoromethyl, a cyano, a nitro, a carboxy, analkoxycarbonyl of 2-7 carbon atoms, an alkanoyl of 2-7 carbon atoms, aphenoxy, a phenyl, a thiophenoxy, a benzoyl, a benzyl, an amino, analkylamino of 1-6 carbon atoms, a dialkylamino of 2 to 12 carbon atoms,a phenylamino, a benzylamino, an alkanoylamino of 1-6 carbon atoms, analkenoylamino of 3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms,a carboxyalkyl of 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbonatoms, an aminoalkyl of 1-5 carbon atoms, an N-alkylaminoalkyl of 2-9carbon atoms, a N,N-dialkylaminoalkyl of 3-10 carbon atoms, anN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, a mercapto, a methylmercapto, an alkanoyloxy of 1-6 carbonatoms, an alkenoyloxy of 3-8 carbon atoms, an alkynoyloxy of 3-8 carbonatoms, a carbamoyl, an N-alkylcarbamoyl of 2-7 carbon atoms, aN,N-dialkylcarbamoyl of 3-13 carbon atoms, and a benzoylamino;a radical of the form:

wherein;A′ is a pyridinyl, a pyrimidinyl, or a phenyl ring; wherein thepyridinyl, pyrimidinyl, or phenyl ring may be optionally mono- ordi-substituted with a substituent comprising an alkyl of 1-6 carbonatoms, an alkenyl of 2-6 carbon atoms, an alkynyl of 2-6 carbon atoms,azido, a hydroxyalkyl of 1-6 carbon atoms, a halogen, a halomethyl, analkoxymethyl of 2-7 carbon atoms, an alkanoyloxymethyl of 2-7 carbonatoms, an alkoxy of 1-6 carbon atoms, an alkylthio of 1-6 carbon atoms,hydroxy, a trifluoromethyl, a cyano, a nitro, a carboxy, analkoxycarbonyl of 2-7 carbon atoms, an alkanoyl of 2-7 carbon atoms, aphenoxy, a phenyl, a thiophenoxy, a benzoyl, a benzyl, an amino, analkylamino of 1-6 carbon atoms, a dialkylamino of 2 to 12 carbon atoms,a phenylamino, a benzylamino, an alkanoylamino of 1-6 carbon atoms, analkenoylamino of 3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms,a carboxyalkyl of 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbonatoms, an aminoalkyl of 1-5 carbon atoms, a N-alkylaminoalkyl of 2-9carbon atoms, a N,N-dialkylaminoalkyl of 3-10 carbon atoms, anN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, a mercapto, a methylmercapto, an alkanoyloxy of 1-6 carbonatoms, a alkenoyloxy of 3-8 carbon atoms, an alkynoyloxy of 3-8 carbonatoms, a carbamoyl, a N-alkylcarbamoyl of 2-7 carbon atoms, aN,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino;T is substituted at a carbon of the pyridinyl, pyrimidinyl, or phenylring with —NH(CH₂)_(m)—, —O(CH₂)_(m)—, —S(CH₂)_(m)—, —NR(CH₂)_(m)—,—(CH₂)_(m)—, —(CH₂)_(m)NH—, —(CH₂)_(m)O—, —(CH₂)_(m)S—, —SO(CH₂)_(m)—,—SO₂(CH₂)_(m)—, —CO(CH₂)_(m)—, —(CH₂)_(m)CO—, —(CH₂)_(m)SO—,—(CH₂)_(m)SO₂— or —(CH₂)_(m)NR—;L is an imidazole or a phenyl ring wherein the imidazole or phenyl ringare optionally substituted at a carbon or nitrogen with one, two, orthree substituents comprising an alkyl of 1-6 carbon atoms, an alkenylof 2-6 carbon atoms, an alkynyl of 2-6 carbon atoms, azido, ahydroxyalkyl of 1-6 carbon atoms, a halogen, a halomethyl, analkoxymethyl of 2-7 carbon atoms, an alkanoyloxymethyl of 2-7 carbonatoms, an alkoxy of 1-6 carbon atoms, an alkylthio of 1-6 carbon atoms,a hydroxy, a trifluoromethyl, a cyano, a nitro, a carboxy, analkoxycarbonyl of 2-7 carbon atoms, an alkanoyl of 2-7 carbon atoms, aphenoxy, a phenyl, a thiophenoxy, a benzoyl, a benzyl, an amino, analkylamino of 1-6 carbon atoms, a dialkylamino of 2 to 12 carbon atoms,a phenylamino, a benzylamino, an alkanoylamino of 1-6 carbon atoms, analkenoylamino of 3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms,a carboxyalkyl of 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbonatoms, an aminoalkyl of 1-5 carbon atoms, an N-alkylaminoalkyl of 2-9carbon atoms, an N,N-dialkylaminoalkyl of 3-10 carbon atoms, aN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, a mercapto, a methylmercapto, an alkanoyloxy of 1-6 carbonatoms, an alkenoyloxy of 3-8 carbon atoms, an alkynoyloxy of 3-8 carbonatoms, a carbamoyl, an N-alkylcarbamoyl of 2-7 carbon atoms, aN,N-dialkylcarbamoyl of 3-13 carbon atoms, a benzoylamino, a 5- or6-membered heteroaryl ring where the heteroaryl ring contains 1 to 3heteroatoms selected from N, O, and S and where the heteroaryl ring maybe optionally mono- or di-substituted with a substituent comprising ahalogen, an oxo, a thiocarbonyl, an alkyl of 1-6 carbon atoms, analkenyl of 2-6 carbon atoms, an alkynyl of 2-6 carbon atoms, azido, ahydroxyalkyl of 1-6 carbon atoms, a halomethyl, an alkoxymethyl of 2-7carbon atoms, an alkanoyloxymethyl of 2-7 carbon atoms, an alkoxy of 1-6carbon atoms, an alkylthio of 1-6 carbon atoms, a hydroxy, atrifluoromethyl, a cyano, a nitro, a carboxy, an alkoxycarbonyl of 2-7carbon atoms, an alkanoyl of 2-7 carbon atoms, a phenoxy, a phenyl, athiophenoxy, a benzoyl, a benzyl, an amino, an alkylamino of 1-6 carbonatoms, a dialkylamino of 2 to 12 carbon atoms, a phenylamino, abenzylamino, an alkanoylamino of 1-6 carbon atoms, an alkenoylamino of3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms, a carboxyalkylof 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbon atoms, anaminoalkyl of 1-5 carbon atoms, an N-alkylaminoalkyl of 2-9 carbonatoms, a N,N-dialkylaminoalkyl of 3-10 carbon atoms, aN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, a mercapto, a methylmercapto, an alkanoyloxy of 1-6 carbonatoms, an alkenoyloxy of 3-8 carbon atoms, an alkynoyloxy of 3-8 carbonatoms, a carbamoyl, an N-alkylcarbamoyl of 2-7 carbon atoms, aN,N-dialkylcarbamoyl of 3-13 carbon atoms, and a benzoylamino;m is 0-3;n is 0-1;

X is NH, O, S, or NR;

R is alkyl of 1-6 carbon atoms;R₁, R₂, R₃ and R₄ are a hydrogen, a halogen, a hydroxy, an amino, ahydroxyamino, a trifluoromethyl, a trifluoromethoxy, a mercapto, analkyl of 1-6 carbon atoms, a cycloalkyl of 3-8 carbon atoms, an alkenylof 2-6 carbon atoms, an alkynyl of 2-6 carbon atoms, an alkenyloxy of2-6 carbon atoms, an alkynyloxy of 2-6 carbon atoms, a hydroxyalkyl of1-6 carbon atoms, a mercaptoalkyl of 1-6 carbon atoms, a halomethyl, analkoxymethyl of 2-7 carbon atoms, an alkoxy of 1-6 carbon atoms, a2-methoxyethoxy, a 2-(2-methoxyethoxy)ethoxy, a cycloalkoxy of 3-8carbon atoms, an alkylthio of 1-6 carbon atoms, a cycloalkylthio of 3-8carbon atoms, an alkysulfinyl of 1-6 carbon atoms, an alkylsulfonyl of1-6 carbon atoms, an alkylsulfonamido of 1-6 carbon atoms, analkenylsulfonamido of 2-6 carbon atoms, an alkynylsulfonamido of 2-6carbon atoms, an alkylcarboxamido of 2-7 carbon atoms, a(N-alkyl)alkylcarboxamido of 3-13 carbon atoms, an alkenylcarboxamido of3-7 carbon atoms, an (N-alkyl)alkenylcarboxamido of 4-13 carbon atoms,an alkynylcarboxamido of 3-7 carbon atoms, an(N-alkyl)alkynylcarboxamido of 4-13 carbon atoms, cyano, nitro, carboxy,a alkoxycarbonyl of 2-7 carbon atoms, an alkanoyl of 2-7 carbon atoms,an alkenoyl of 3-7 carbon atoms, a N-alkyl-N-alkenylamino of 4 to 12carbon atoms, an N,N-dialkenylamino of 6-12 carbon atoms, phenylamino,benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, an alkylamino of 1-6carbon atoms, an alkanoyloxy of 2-7 carbon atoms, an alkenoyloxy of 3-8carbon atoms, an alkynoyloxy of 3-8 carbon atoms, a carbamoyl, anN-alkylcarbamoyl of 2-7 carbon atoms, a N,N-dialkylcarbamoyl of 3-13carbon atoms, a dialkylamino of 2 to 12 carbon atoms, analkanoyloxymethyl group of 2-7 carbon atoms, an alkenoyloxymethy groupof 2-7 carbon atoms, an alkynoyloxymetho group of 2-7 carbon atoms,azido, benzoyl, a carboxyalkyl of 2-7 carbons, and a carboalkoxyalkyl of3-8 carbon atoms,

—NR₆C(O)H; —N(C(O)R₆)C(O)R₆;

—Y—(C(R₆)₂)_(p)-Het1;Y—(C(R₆)₂)_(p)-Het1-(C(R₆)₂)_(q)-Z-(C(R₆)₂)_(r)-Het2;Het1 is a 3-8 membered saturated heterocyclic ring containing one ormore nitrogen, oxygen or sulfur atoms such as, but not limited tomorpholine, thiomorpholine, thiomorpholine S-oxide, thiomorpholineS,S-dioxide, piperidine, pyrrolidine, aziridine, piperazine,tetrahydrothiophene, tetrahydrofuran, dioxane, 1,3-dioxolane pyrrole,tetrahydropyran, and diazepan; wherein Het1 is optionally mono- ordi-substituted on a carbon or a nitrogen with R₆; optionally mono- ordi-substituted on a carbon with hydroxy, —N(R₆)₂, or —OR₆; optionallymono or di-substituted on carbon with the mono-valent radicals—(C(R₆)₂)_(s)OR₆ or —[(C(R₆)₂)_(s)N(R₆)₂]; or optionally mono ordi-substituted on a saturated carbon with divalent radicals ═O or—O(C(R₆)₂)_(s)O—;Het2 is a heteroaryl selected from the group comprising morpholine,thiomorpholine, thiomorpholine S-oxide, thiomorpholine S,S-dioxide,piperidine, pyrrolidine, aziridine, pyridine, imidazole, 1,2,3-triazole,1,2,4-triazole, thiazole, thiazolidine, tetrazole, piperazine, furan,thiophene, tetrahydrothiophene, tetrahydrofuran, dioxane, 1,3-dioxolanepyrrole, and tetrahydropyran; wherein Het2 is optionally mono- ordi-substituted on carbon or nitrogen with R₆; optionally mono- ordi-substituted on carbon with hydroxy, —N(R₆)₂, or —OR₆; optionally monoor di-substituted on carbon with the mono-valent radicals—(C(R₆)₂)_(s)OR₆ or —[(C(R₆)₂)_(s)N(R₆)₂]; or optionally mono ordi-substituted on a saturated carbon with divalent radicals ═O or—O(C(R₆)₂)_(s)O—;R₆ is hydrogen, an alkyl of 1-6 carbon atoms, an alkenyl of 2-6 carbonatoms, an alkynyl of 2-6 carbon atoms, a cycloalkyl of 1-6 carbon atoms,an alkanoyl of 2-7 carbon atoms, a carbamoylalkyl of 2-7 carbon atoms, ahydroxyalkyl of 1-6 carbon atoms, a hydroxycycloalkyl of 3-6 carbonatoms, a carboxyalkyl of 2-7 carbon atoms, pyrrolidine, piperidine, orimidazole optionally substituted with methyl;a phenyl optionally mono-, di-, or tri-substituted with halogen, analkoxy of 1-6 carbon atoms, trifluoromethyl, amino, an alkylamino of 1-3carbon atoms, a dialkylamino of 2-6 carbon atoms, nitro, cyano, azido,halomethyl, an alkoxymethyl of 2-7 carbon atoms, an alkanoyloxymethyl of2-7 carbon atoms, an alkylthio of 1-6 carbon atoms, hydroxy, a carboxyl,a alkoxycarbonyl of 2-7 carbon atoms, phenoxy, phenyl, thiophenoxy,benzoyl, benzyl, phenylamino, benzylamino, an alkanoylamino of 1-6carbon atoms or alkyl of 1-6 carbon atoms;Y is O, S, —NR₆C(O)—, —C(O)NR₆—, NR₆ or a bond;Z is O, S, —NR₆C(O)—, —C(O)NR₆—, NR₆ or a bond;p is 0-4;q is 0-4;r is 0-4;s is 1-6; andat least one of the groups, R₁, R₂, R₃ or R₄ is consisting of—Y—(C(R₆)₂)_(p)-Het1-(C(R₆)₂)_(q)-Z-(C(R₆)₂)_(r)-Het2;X′ is fluoro or chloro provided that when X′ is chloro, R₂ is nitro;G′ is bromo, iodo, or sulfonate;E is a leaving group or a sulfonate;Q is a leaving group, aldehyde, and a ketone;G is hydrogen, a halogen, hydroxy, amino, hydroxyamino, trifluoromethyl,trifluoromethoxy, mercapto, an alkyl of 1-6 carbon atoms, a cycloalkylof 3-8 carbon atoms, an alkenyl of 2-6 carbon atoms, a alkynyl of 2-6carbon atoms, a alkenyloxy of 2-6 carbon atoms, an alkynyloxy of 2-6carbon atoms, a hydroxyalkyl of 1-6 carbon atoms, a mercaptoalkyl of 1-6carbon atoms, halomethyl, an alkoxymethyl of 2-7 carbon atoms, an alkoxyof 1-6 carbon atoms, a 2-methoxyethoxy, a 2-(2-methoxyethoxy)ethoxy, acycloalkoxy of 3-8 carbon atoms, an alkylthio of 1-6 carbon atoms, acycloalkylthio of 3-8 carbon atoms, an alkysulfinyl of 1-6 carbon atoms,an alkylsulfonyl of 1-6 carbon atoms, an alkylsulfonamido of 1-6 carbonatoms, an alkenylsulfonamido of 2-6 carbon atoms, an alkynylsulfonamidoof 2-6 carbon atoms, an alkylcarboxamido of 2-7 carbon atoms, a(N-alkyl)alkylcarboxamido of 3-13 carbon atoms, an alkenylcarboxamido of3-7 carbon atoms, an (N-alkyl)alkenylcarboxamido of 4-13 carbon atoms,an alkynylcarboxamido of 3-7 carbon atoms, an(N-alkyl)alkynylcarboxamido of 4-13 carbon atoms, cyano, nitro, carboxy,a alkoxycarbonyl of 2-7 carbon atoms, an alkanoyl of 2-7 carbon atoms,an alkenoyl of 3-7 carbon atoms, a N-alkyl-N-alkenylamino of 4 to 12carbon atoms, an N,N-dialkenylamino of 6-12 carbon atoms, phenylamino,benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, an alkylamino of 1-6carbon atoms, an alkanoyloxy of 2-7 carbon atoms, an alkenoyloxy of 3-8carbon atoms, an alkynoyloxy of 3-8 carbon atoms, a carbamoyl, anN-alkylcarbamoyl of 2-7 carbon atoms, a N,N-dialkylcarbamoyl of 3-13carbon atoms, a dialkylamino of 2 to 12 carbon atoms, analkanoyloxymethyl group of 2-7 carbon atoms, an alkenoyloxymethyl groupof 2-7 carbon atoms, an alkynoyloxymethyl group of 2-7 carbon atoms,azido, benzoyl, a carboxyalkyl of 2-7 carbons, a carboalkoxyalkyl of 3-8carbon atoms,

—NR₆C(O)H; —N(C(O)R₆)C(O)R₆; or

a crystalline form or a pharmaceutically acceptable salt thereof.

Preferred compounds of the invention include the compounds of Formula 1as follows:

-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-(4-methyl    piperazin-1-yl)quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino-6-methoxy-7-piperazin-1-ylquinoline-3-carbonitrile;-   6-methoxy-4-{[4-(2-methoxyphenoxy)phenyl]amino}-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile,    trifluoroacetate salt;-   6-methoxy-4-[4-(pyridin-2-ylsulfanyl)-phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)-quinoline-3-carbonitrile;-   4-[3-chloro-4-(pyridin-2-ylsulfanyl)-phenylamino]-6-methoxy-7-(4-pyrrolidin-1-yl-piperidin-1-yl)-quinoline-3-carbonitrile;-   6-methoxy-4-[4-(pyridin-3-yloxy)-phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)-quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-(2-methoxyethoxy)-7-(4-pyrrolidin-1-ylpiperidin-1-yl))quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-ethoxy-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile;-   7-(1,4′-bipiperidin-1′-yl)-4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxyquinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-(4-morpholin-4-ylpiperidin-1-yl)quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-[4-(1-methylpiperidin-4-yl)piperazin-1-yl]quinoline-3-carbonitrile;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-{4-[2-(1H-imidazole-1-yl)ethyl]piperazin-1-yl)-6-methoxyquinoline-3-carbonitrile;-   6-Nitro-4-oxo-7-(4-pyrrolidin-1-ylpiperidin-1-yl)-1,4-dihydroquinoline-3-carbonitrile;-   4-chloro-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile,-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile;-   6-amino-4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamino]-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile;-   N-acetyl-N-[4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-3-cyano-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-6-yl]acetamide;-   N-[4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-3-cyano-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-6-yl]acetamide;-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-(methylamino)-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile;-   4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile;-   7-(2-[1,4′]bipiperidinyl-1′-yl-ethoxy)-6-methoxy-4-(4-phenoxy-phenylamino)-quinoline-3-carbonitrile;-   4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrile;-   4-(2,4-dichloro-5-methoxyanilino)-6-methoxy-7-{3-[4-(1-pyrrolidinyl)-1-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrile;-   4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{2-[4-(1-pyrrolidinyl)-1-piperidinyl]ethoxy}-3-quinolinecarbonitrile;-   6-methoxy-4-({4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-(4-pyrrolidin-1-yl    piperidin-1-yl)quinoline-3-carbonitrile; and-   4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-hydroxy-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile.

In another embodiment this invention provides a method of inhibiting aRas/Raf/MEK signaling cascade in a mammal comprising administering tothe mammal an effective amount of a compound of Formula 1:

wherein the substitutions on the ring are previously described andincludes a crystalline form or a pharmaceutically acceptable saltthereof.

In one embodiment the inhibition of the Ras/Raf/MEK signaling cascadeprevents or inhibits stroke, osteoporosis, cancer, rheumatoid arthritisand inflammatory disease, polycystic kidney disease, and colonic polyps.

In a preferred embodiment the cancer is a melanoma, a pancreatic cancer,or a lung cancer.

In another embodiment the Ras/Raf/MEK signaling cascade in a mammal isinhibited by providing the mammal with an effective amount of at leastone pharmaceutical composition, wherein the at least one pharmaceuticalcomposition includes a compound of Formula 1:

wherein the substitutions on the ring are previously described andincludes a crystalline form or a pharmaceutically acceptable saltthereof.

In a preferred embodiment the mammal is provided with an effectiveamount of at least one biologically active agent provided prior to theat least one pharmaceutical composition, concurrently with the at leastone pharmaceutical composition or after the at least one pharmaceuticalcomposition.

An embodiment of this invention includes a means for preparing acompound of Formula I comprising:

a. reacting a compound of Formula 2

with a heated compound of Formula 3

Het2(C(R₆)₂)_(r)Z-(C(R₆)₂)_(q)-Het1(C(R₆)₂)_(p)—YH  3

optionally with a base;

b. to yield the compound of Formula 1 having the structure

wherein the substitutions on the ring are previously described.

In another embodiment the invention includes a means for preparing acompound of Formula I comprising:

a. reacting a compound of Formula 2 or 7

with a heated compound of Formula 5

optionally with a base;

b. to yield the compound of Formula 1 having the structure

wherein the substitutions on the ring are previously described.

In another embodiment the invention describes a means for preparing acompound of Formula I comprising:

a. combining a compound of formula 9

with a compound of Formula 10

Het2(C(R₆)₂)_(r)-ZH

to obtain a compound of Formula 1

wherein the substitutions on the ring are previously described.

In an embodiment this invention includes a means for preparing acompound of Formula I comprising

a. combining a compound of Formula 12 or 17

with a compound of Formula 3

Het2(C(R₆)₂)_(r)-Z-(C(R₆)₂)_(q)-Het1(C(R₆)₂)_(p)—YH  3

wherein, when G′ is present a palladium catalyst is requiredto obtain a compound of Formula 13 or 18

b. combining the compound of Formula 13 or 18 with a halogenating agentin the presence of PO(Z)₃ to obtain a compound of Formula 14 or 19

c. combining the compound of Formula 14 and 19 with a compound ofFormula 15

to obtain a compound of Formula 1 or Formula 11

wherein the substitutions on the ring are previously described.

In another embodiment the invention includes a compound of Formula 12

wherein the substitutions on the ring are previously described.

In another embodiment the invention includes a compound of Formula 13

wherein the substitutions on the ring are previously described.

An embodiment the invention includes a compound of formula 18

wherein the substitutions on the ring are previously described.

In an embodiment this invention includes a compound of Formula 19

wherein the substitutions on the ring are previously described.

In an embodiment this invention includes a compound of Formula 7

wherein the substitutions on the ring are previously described.

In another embodiment this invention includes a compound of formula 9

wherein the substitutions on the ring are previously described.

The following experimental details are set forth to aid in anunderstanding of the invention, and are not intended, and should not beconstrued, to limit in any way the invention set forth in the claimsthat follow thereafter.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with this invention there is provided a group of compoundsrepresented by Formula 1:

wherein the substitutions on the ring are previously described herein.

For purposes of this invention various terms are defined in thefollowing paragraphs.

Alkanoyl is defined as a —COR″ radical, where R″ is an alkyl radical of1-6 carbon atoms. Alkenoyl is defined as a —COR″ radical, where R″ is analkenyl radical of 2-6 carbon atoms. Alkanoyloxy is defined as a —OCOR″radical, where R″ is an alkyl radical of 1-6 carbon atoms.Alkanoyloxymethyl is defined as R″CO₂CH₂— radical, where R″ is an alkylradical of 1-6 carbon atoms.

The terms “alkenyl” and “alkynyl” refer to unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but which contain at least one double or triple carbon-carbonbond, respectively, in all possible configurational isomers, for examplecis and trans. The alkenyl portion of the alkenyl, alkenyloxy,alkenylsulfonamido, substituents include both straight chain as well asbranched carbon chains and one or more sites of unsaturation and allpossible configurational isomers.

The term “alkoxy” as used herein refers to an alkyl group, as definedabove, having an oxygen radical attached thereto. Representative alkoxylgroups include methoxy, ethoxy, propyloxy, tert-butoxy and the like. Theterm alkoxy can be used alone or as part of a chemical name as in forexample, “alkoxy-enaminonitrile”. The terms “alkoxyl” or “alkoxy” asused herein refers to an alkyl group, as defined above, having an oxygenradical attached thereto. Representative alkoxyl groups include methoxy,ethoxy, propyloxy, tert-butoxy and the like. Alkoxycarbonyl of 2-7carbon atoms is defined as a —CO₂R″ radical, where R″ is an alkylradical of 1-6 carbon atoms.

Akoxymethyl is defined as R″OCH₂ radical where R″ is an alkyl radical of1-6 carbon atoms.

The term “alkyl” refers to the radical of saturated aliphatic groups,including straight-chain alkyl groups, branched-chain alkyl groups,cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, andcycloalkyl substituted alkyl groups. In a preferred embodiment, astraight chain or branched chain alkyl has 6 or fewer carbon atoms inits backbone. The term “alkyl” can be used alone or as part of achemical name as in for example, alkoxy, alkanoyloxy, alkoxymethyl,alkanoyloxymethyl, alkysulfinyl, alkylsulfonyl, alkylsulfonamido,alkoxycarbonyl, alkynoyloxy, alkanoylamino, N-alkylcarbamoyl,N,N-dialkylcarbamoyl, N-alkylaminoalkoxy, N-alkyl-amino-alkyl,N,N-dialkylaminoalkyl, hydroxyalkyl, alkylthio, andN,N-dialkylaminoalkoxy include both straight chain as well as branchedcarbon chains.

Alkylcarboxamido, alkenylcarboxamido, alkynylcarboxamido are defined asR″CONH— radical, where R″ is an alkyl radical of 1-6 carbon atoms, analkenyl radical of 2-6 carbon atoms, or an alkynyl radical of 2-6 carbonatoms, respectively. (N-alkyl)alkenylcarboxamido, alkynylcarboxamido,(N-alkyl)alkynylcarboxamido are defined as R″CO(R′)N— radical, where R′is an alkyl radical of 1-6 carbon atoms, R″ is an alkyl radical of 1-6carbon atoms, an alkenyl radical of 2-6 carbon atoms, or an alkynylradical of 2-6 carbon atoms, respectively. N-alkylcarbamoyl is definedas R″NHCO— radical, where R″ is an alkyl radical of 1-6 carbon atoms.N,N-dialkylcarbamoyl is defined as R″R′NCO— radical, where R″ is analkyl radical of 1-6 carbon atoms, R′ is an alkyl radical of 1-6 carbonatoms and R′ and R″ may be the same or different.

Alkysulfinyl is defined as R″SO— radical, where R″ is an alkyl radicalof 1-6 carbon atoms. Alkylsulfonyl is defined as R″SO₂— radical, whereR″ is an alkyl radical of 1-6 carbon atoms. Alkylsulfonamido,alkenylsulfonamido, alkynylsulfonamido are defined as R″SO₂NH— radical,where R″ is an alkyl radical of 1-6 carbon atoms, an alkenyl radical of2-6 carbon atoms, or an alkynyl radical of 2-6 carbon atoms,respectively.

The term “alkylthio” refers to an alkyl group, as defined above, havinga sulfur radical attached thereto. In preferred embodiments, the“alkylthio” moiety is represented by one of —S-alkyl, —S-alkenyl,—S-alkynyl, and —S—. Representative alkylthio groups include methylthio,ethylthio, and the like.

The terms “amine”, “amino”, and “amide” are art recognized and refer toboth unsubstituted and substituted amines. The terms can be used incombination with other terms described herein, for example, alkylamino,aminoalkyl, dialkylamino, alkanoylamino, and alkenoylamino.

The term “aryl” as used herein includes 4-, 5-, 6-, 7- and 10-memberedsingle ring or fused multiple rings aromatic groups, which maysubstituted or unsubstituted. For purposes of this invention the term“aryl” is defined as an aromatic carbocyclic moiety and may besubstituted or unsubstituted. Preferred aryl groups have 6 to 14 carbonatoms. Particularly preferred aryl groups are phenyl and napthyl. Thearomatic ring can be optionally independently mono-, di-, tri- ortetra-substituted. Preferred substituents are selected from the groupconsisting of, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl,cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromaticor heteroaromatic moieties and —CN. The term “aryl” also includespolycyclic ring systems having two or more rings in which two or morecarbons are common to two adjoining rings (the rings are “fused”)wherein at least one of the carbocyclic rings is aromatic, e.g., theother rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, and/oraryls.

The term azido represents a compound that includes a group N:N.N—.

For purposes of this invention a base could be, but is not limited to,triethylamine, Huenig's base (diisopropylethylamine), or an inorganicbase such as potassium carbonate.

Benzoyl is a term that represents a benzene with a carbonyl with theattachment at the carbonyl. Such term can be used alone or incombination with terms previously describe, as for example,benzoylamino.

The term carbamoyl represents the radical NH₂CO— from carbamic acid.

The term carbonyl represents the radical ═CO.

Carboxy is defined as a —CO₂H radical. Carboxyalkyl is defined as aHO₂C—R′″— radical where R′″ is a divalent alkyl radical of 1-6 carbonatoms, Carboalkoxyalkyl is defined as a R″O₂C—R′″— radical where R′″ isa divalent alkyl radical and where R″ and R′″ may be the same ordifferent, and together have 2-7 carbon atoms.

Cyano represents the radical NC—.

The cycloalkyl portions of cycloalkyl, N-cycloalkylamino, N-cycloalkyl-Nalkylaminoalkyl, N,N-dicycloalkylaminoalkyl, cycloalkylthio andazacycloalkyl substituents include both simple carbocycles as well ascarbocycles containing alkyl substituents, where a carbocycle is acyclic compound where all the ring members are carbon.

The compounds of this invention may contain one or more asymmetriccarbons atoms; in such cases, the compounds of this invention includethe individual diasteromers, the racemates, and the individual R and Sentantiomers thereof. Some of the compounds of this invention maycontain one or more double bonds; in such cases, the compounds of thisinvention include each of the possible configurational isomers as wellas mixtures of these isomers. The compounds of this invention maycontain an asymmetric carbon atom and some of the compounds of thisinvention may contain one or more asymmetric centers and may thus giverise to stereoisomers, such as enantiomers and diastereomers. Thestereoisomers of the instant invention are named according to theCahn-Ingold-Prelog System. While shown without respect tostereochemistry in Formula (I), the present invention includes all theindividual possible stereoisomers; as well as the racemic mixtures andother mixtures of R and S stereoisomers (scalemic mixtures which aremixtures of unequal amounts of enantiomers) and pharmaceuticallyacceptable salts thereof. It should be noted that stereoisomers of theinvention having the same relative configuration at a chiral center maynevertheless have different R and S designations depending on thesubstitution at the indicated chiral center.

The term “halogen” refers to an atom of fluorine, chlorine, bromine, oriodine. A halogen can be combined with halogens or other groups, as forexample in the case of, halomethyl or trifluoromethyl.

The term “heteroaryl” refers to a 3 to 8 membered ring structure, whichring structure includes one to four heteroatoms. Heteroaryls include,but are not limited to, pyrrolidine oxolane, thiolane, piperidine,piperazine, pyrrole, furan, thiophene, imidazole, oxazole, thiazole,triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, andmorpholine. For purposes of this invention a heteroaryl comprises aheterocyclic ring system of one to three fused rings and contains 1 to 4heteroatoms the same or different selected from the group consisting ofS, N, and O. The remaining rings of the ring system may be fullyunsaturated, partially saturated, or fully saturated. Each ringcomprises three to ten members. Preferred heteroaryl groups arethiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene,phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole,pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole,indole, indazole, purine, quinolizine, isoquinoline, quinoline,phthalazine, naphthyridine, quinoxaline, quinazoline, pteridine,carbazole, phenanthridine, acridine, perimidine, phenanthroline,phenazine, phenothiazine, furazan, phenoxazine and pyrrolidine. Theheteroaryl can be independently substituted at one or more positions.Preferred substituents are halogen, alkyl, aralkyl, alkenyl, alkynyl,cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, acyl, aldehyde, ester, a cycloheteroalkyl, an aromatic orheteroaromatic moiety, —CN, or Y. When the heteroaryl is substitutedwith Y, wherein Y is —NH, —O—, —S—, or —NR—, wherein R is an alkyl of1-6 carbon atoms, at one position on the ring there is furthersubstitution on the —NH, —O—, —S—, or —NR— with a (CH₂)n-X group. Forpurposes of this invention n is 0-1 and “X” is cycloalkyl of 3 to 7carbon atoms, which may be optionally substituted with one or more alkylof 1 to 6 carbon atoms; or is a pyridinyl, pyrimidinyl, or phenyl ring;wherein the pyridinyl, pyrimidinyl, or phenyl ring may be optionallymono- di-, or tri-substituted with substituents independently selectedfrom the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenylof 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms,alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms,alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro,carboxy, carboalkoxy of 2-7 carbon atoms, carboalkyl of 2-7 carbonatoms, phenoxy, phenyl, thiophenoxy, benzoyl, benzyl, amino, alkylaminoof 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, phenylamino,benzylamino, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8carbon atoms, alkynoylamino of 3-8 carbon atoms, benzoylamino, and-Q-(CH₂)_(m)Ar, wherein Q is selected from O, NH, N(C₁-C₆ alkyl) or S, mis 0, 1 or 2, and Ar is phenyl or pyridyl optionally substituted withone to three moieties independently selected from halogen, alkyl of 1-6carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms,azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl,cyano, nitro, carboxy, carboalkoxy of 2-7 carbon atoms, carboalkyl of2-7 carbon atoms, phenoxy, phenyl, thiophenoxy, benzoyl, benzyl, amino,alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms,phenylamino, benzylamino, alkanoylamino of 1-6 carbon atoms,alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms andbenzoylamino.

It is understood by one skilled in the art that the heteroaryl orbicyclic heteroaryl Ar groups of the compounds of Formula I do notcontain O—O, S—S, or S—O bonds, as they would be unstable. Preferredbicyclic aryl or bicyclic heteroaryl ring systems include naphthalene,tetralin, indan, 1-indanone, 1,2,3,4-tetrahydroquinoline, naphthyridine,benzofuran, 3-oxo-1,3-dihydroisobenzofuran, benzothiophene,1,1-dioxo-benzothiophene, indole, indoline1,3-dioxo-2,3-dihydro-1H-isoindole, benzotriazole, 1H-indazole,indoline, indazole, 1,3-benzodioxole, benzoxazole, purine, phthalimide,coumarin, chromone, quinoline, tetrahydroquinoline, isoquinoline,benzimidazole, quinazoline, pyrido[2,3-b]pyridine,pyrido[3,4-b]pyrazine, pyrido[3,2-c]pyridazine, pyrido[3,4-b]pyridine,1H-pyrazole[3,4-d]pyrimidine, 1,4-benzodioxane, pteridine,2(1H)-quinolone, 1(2H)isoquinolone, 2-oxo-2,3-dihydrobenzthiazole,1,2-methylenedioxybenzene, 2-oxindole, 1,4-benzisoxazine, benzothiazole,quinoxaline, quinoline-N-oxide, isoquinoline-N-oxide,quinoxaline-N-oxide, quinazoline-N-oxide, benzoazine, phthalazine,1,4-dioxo-1,2,3,4-tetrahydrophthalazine, 2-oxo-1,2-dihydro-quinoline,2,4-dioxo-1,4-dihydro-2H-benzo[d][1,3]oxazine,2,5-dioxo-2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepine, or cinnoline.

When L is a 5 or 6-membered heteroaryl ring, preferred heteroaryl ringsare pyridine, pyrimidine, imidazole, thiazole, thiazolidine, pyrrole,furan, thiophene, oxazole, or 1,2,4-triazole.

Either or both rings of the bicyclic aryl or bicyclic heteroaryl Argroup may be fully unsaturated, partially saturated, or fully saturated.An oxo substituent on the bicyclic aryl or bicyclic heteroaryl moietymeans that one of the carbon atoms has a carbonyl group. A thiocarbonylsubstituent on the bicyclic aryl or bicyclic heteroaryl moiety meansthat one of the carbon atoms has a thiocarbonyl group.

When L is a 5 or 6-membered heteroaryl ring, it may be fully unsaturatedor partially saturated. The heteroaryl ring can be bound to A′ viacarbon or nitrogen.

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen,sulfur, phosphorus, and selenium. The term “heteroatom” as used hereinmeans an atom of any element other than carbon or hydrogen. Preferredheteroatoms are nitrogen, oxygen, sulfur and phosphorous.

Mercapto is a prefix or suffix representing a thio group —SH, forexample, methylmercapto.

As used herein, the term “nitro” means —NO₂; the term “halogen”designates —F, —Cl, —Br or —I; the term “sulfhydryl” means —SH; the term“hydroxyl” means —OH; and the term “sulfonyl” means —SO—.

The term phenoxy represents the radical PhO, a form of phenol. A phenolis an aryl hydroxide. Both terms can be used alone or in conjunctionwith terms described herein, such as, thiophenoxy or phenylamino.

The term oxo is a prefix indicating the keto group, as for exampleoxomalonic acid, HOOC—CO—COOH.

The pharmaceutically acceptable salts are those derived from suchorganic and inorganic acids such as: acetic, lactic, citric, tartaric,succinic, maleic, malonic, gluconic, hydrochloric, hydrobromic,phosphoric, nitric, sulfuric, methanesulfonic, and similarly knownacceptable acids.

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents of organic compounds include acyclic andcyclic, branched and unbranched, carbocyclic and heterocyclic, aromaticand nonaromatic substituents of organic compounds. It will be understoodthat “substitution” or “substituted with” includes the implicit provisothat such substitution is in accordance with permitted valence of thesubstituted atom and the substituent, and that the substitution resultsin a stable compound, e.g., which does not spontaneously undergotransformation such as by rearrangement, cyclization, elimination, etc.

The compounds of this invention when part of a pharmaceuticalcomposition, may be formulated with conventional excipients, such as afiller, a disintegrating agent, a binder, a lubricant, a flavoringagent, a color additive, or a carrier. The carrier may be for example adiluent, an aerosol, a topical carrier, an aqueous solution, anonaqueous solution or a solid carrier. The carrier may be a polymer ora toothpaste. A carrier in this invention encompasses any of thestandard pharmaceutically accepted carriers, such as phosphate bufferedsaline solution, acetate buffered saline solution, water, emulsions suchas an oil/water emulsion or a triglyceride emulsion, various types ofwetting agents, tablets, coated tablets and capsules. When providedorally or topically, such compounds would be provided to a subject bydelivery in different carriers. Typically, such carriers containexcipients such as starch, milk, sugar, certain types of clay, gelatin,stearic acid, talc, vegetable fats or oils, gums, or glycols. Thespecific carrier would need to be selected based upon the desired methodof delivery, for example, phosphate buffered saline (PBS) could be usedfor intravenous or systemic delivery and vegetable fats, creams, salves,ointments or gels may be used for topical delivery.

The compounds of this invention when part of a pharmaceuticalcomposition, of the present invention may be delivered together withsuitable diluents, preservatives, solubilizers, emulsifiers, adjuvantsand/or carriers useful in treatment or prevention of neoplasm. Suchcompositions are liquids or lyophilized or otherwise dried formulationsand include diluents of various buffer content (for example, Tris-HCl,acetate, phosphate), pH and ionic strength, additives such as albuminsor gelatin to prevent absorption to surfaces, detergents (for example,TWEEN 20, TWEEN 80, PLURONIC F68, bile acid salts), solubilizing agents(for example, glycerol, polyethylene glycerol), anti-oxidants (forexample ascorbic acid, sodium metabisulfate), preservatives (forexample, thimerosal, benzyl alcohol, parabens), bulking substances ortonicity modifiers (for example, lactose, mannitol), covalent attachmentof polymers such as polyethylene glycol, complexation with metal ions,or incorporation of the compound into or onto particulate preparationsof hydrogels or liposomes, micro-emulsions, micelles, unilamellar ormultilamellar vesicles, erythrocyte ghosts, or spheroblasts. Suchcompositions will influence the physical state, solubility, stability,rate of in vivo release, and rate of in vivo clearance of the compoundor composition. The choice of compositions will depend on the physicaland chemical properties of the compound capable of treating orpreventing a neoplasm.

The compounds of this invention may be provided orally, byintralesional, intraperitoneal, intramuscular or intravenous injection;infusion; liposome-mediated delivery; topical, nasal, anal, vaginal,sublingual, uretheral, transdermal, intrathecal, ocular or oticdelivery. In order to obtain consistency in providing the compound ofthis invention it is preferred that a compound of the invention is inthe form of a unit dose. Suitable unit dose forms include tablets,capsules and powders in sachets or vials. Such unit dose forms maycontain from 0.1 to 300 mg of a compound of the invention and preferablyfrom 2 to 100 mg. Still further preferred unit dosage forms contain 5 to50 mg of a compound of the present invention. The compounds of thepresent invention can be administered orally at a dose range of about0-01 to 100 mg/kg or preferably at a dose range of 0.1 to 10 mg/kg. Suchcompounds may be administered from 1 to 6 times a day, more usually from1 to 4 times a day. The effective amount will be known to one of skillin the art; it will also be dependent upon the form of the compound. Oneof skill in the art could routinely perform empirical activity tests todetermine the bioactivity of the compound in bioassays and thusdetermine what dosage to administer. The compound of the presentinvention may be delivered locally via a capsule that allows a sustainedrelease of the compound over a period of time. Controlled or sustainedrelease compositions include formulation in lipophilic depots (forexample, fatty acids, waxes, oils).

The dose provided to a patient will vary depending upon what is beingprovided, the purpose of providing, the manner of providing, and thelike. A “therapeutically effective amount” is an amount sufficient toinhibit an Ras/Raf/MEK signaling cascade in a mammal comprising. Theeffective dosage of active ingredient employed may vary depending on theparticular compound employed, the mode of administration and theseverity of the condition being treated. However, in general,satisfactory results are obtained when the compounds of the inventionare administered at a daily dosage of from about 0.5 to about 1000 mg/kgof animal body weight, optionally given in divided doses two to fourtimes a day, or in sustained release form. For most large mammals thetotal daily dosage is from about 1 to 1000 mg, preferably from about 2to 500 mg. Dosage forms suitable for internal use comprise from about0.5 to 1000 mg of the active compound in intimate admixture with a solidor liquid pharmaceutically acceptable carrier. This dosage regimen maybe adjusted to provide the optimal therapeutic response. For example,several divided doses may be administered daily or the dose may beproportionally reduced as indicated by the exigencies of the therapeuticsituation.

For purposes of this invention a “neoplasm” is defined as cells selectedfrom the breast, kidney, bladder, mouth, larynx, esophagus, stomach,colon, ovary, pancreas, brain, prostrate and lung having a morphologynot found in the majority of the cells of a mammal.

As used in accordance with this invention, the term providing aneffective amount of a compound means either directly administering suchcompound, or administering a prodrug, derivative, or analog which willform an effective amount of the compound within the body.

In one embodiment, the present invention provides for a method ofinhibiting the neoplasm. The method comprises contacting a cell with anamount of a compound effective to decrease or prevent Ras/Raf/MEKsignaling cascade function. The cell may be a mammalian cell and morespecifically a human cell. The cell may also be a bacterial cell such asfor example E coli. The cell may include but is not limited to, aneuronal cell, an endothelial cell, a glial cell, a microglial cell, asmooth muscle cell, a somatic cell, a bone marrow cell, a liver cell, anintestinal cell, a germ cell, a myocyte, a mononuclear phagocyte, anendothelial cell, a tumor cell, a lymphocyte cell, a mesangial cell, aretinal epithelial cell, a retinal vascular cell, a ganglion cell or astem cell. The cell may be a normal cell, an activated cell, aneoplastic cell, a diseased cell, or an infected cell.

In another embodiment, the present invention provides a method for thetreatment or prevention of a neoplasm in a mammal. The present inventionaccordingly provides to a mammal, a pharmaceutical composition thatcomprises a compound of this invention in combination or associationwith a pharmaceutically acceptable carrier. The compound of thisinvention may be administered alone or in combination with othertherapeutically effective compounds or therapies for the treatment orprevention of the neoplasm.

The present invention further provides a compound of the invention foruse as an active therapeutic substance for preventing neoplasm.

The present invention further provides a method of treating neoplasm inhumans, which comprises administering to the infected individual aneffective amount of a compound or a pharmaceutical composition of theinvention.

The present invention provides a method of inhibiting Ras/Raf/MEKsignaling cascade. The method comprises contacting a cell with an amountof a compound effective to decrease or prevent the Ras/Raf/MEK signalingcascade. The cell may be a mammalian cell and more specifically a humancell. The cell may be a normal cell, an activated cell, a neoplasticcell, a diseased cell, or an infected cell. The cell is not limited to acell type.

As used herein the at least one biologically active agent is selectedfrom the group consisting of interferon, a pegylated interferon, aribavirin, protease inhibitors, polymerase inhibitors, small interferingRNA compounds, anti-sense compounds, nucleotide analogs, nucleosideanalogs, immunoglobulins, immunomodulators, hepatoprotectants,anti-inflammatory agents, antibiotics, antivirals, and anti-infectivecompounds.

The preparation of the compounds and intermediates of this inventionencompassed by Formula 1 is described below in Flowsheet 1 wherein Ar,X, n, p, q, r, R₂, R₆, Z, Het1 and Het2 are herein before defined. Y isO, S or NR₆.

Intermediates 2, wherein Ar, X, n and R₂ are herein before defined, andX′ is fluoro or chloro (with the proviso that when X′ is chloro, R₂ isnitro) can be prepared by methods outlined in U.S. Pat. Nos. 6,002,008,6,288,082, and 6,297,258, hereby incorporated by reference.Intermediates 3, wherein p, q, r, R₆, Z, Het1 and Het2 are herein beforedefined, and Y is O, S or NR₆, are commercially available, or can beprepared by methods in the chemistry literature by a chemist skilled inthe art. Displacement of the flourine atom of compounds of Formula 2 atC-7 can be carried out by heating with compounds of Formula 3, in thepresence or absence of added base to provide target compounds of Formula1.

The preparation of compounds of Formula 6 can be carried out as outlinedin Flowsheet 2. Nucleophile 5, wherein Het1 possesses a reactive NHmoiety, q, r, R₆, Z and Het2 are herein before defined, can be reactedwith intermediate 2, wherein Ar, X, X′, n and R₂ are herein beforedefined, at elevated temperatures, with or without added base to providecompounds of Formula 6.

The preparation of compounds of Formula 20 can be carried out asoutlined in Flowsheet 3. Compounds of Formula 7, wherein Ar, X, n, p, R₆are herein before defined, E is a leaving group such as, but notrestricted to halo, tosylate, mesylate, Y is O, G is R₁, R₂ or R₄, someof which can be prepared as described in U.S. Pat. Nos. 6,288,082 and6,297,258, hereby incorporated by reference. Nucleophile 5, wherein Het1possesses a reactive NH moiety, q, r, R₆, Z and Het2 are herein beforedefined, can be reacted with intermediate 7 to provide compounds ofFormula 6 using chemistry methodology described in U.S. Pat. Nos.6,288,082 and 6,297,258, hereby incorporated by reference.

The preparation of the compounds and intermediates of this inventionencompassed by Formula 11 can also be carried out by methods outlined inFlowsheet 4 wherein Ar, X, n, p, q, r, R₆, Z, Y, G, Het1 and Het2 areherein before defined.

Intermediates 9, wherein Ar, X, Y, Het1, R₆, p, q, n and G are hereinbefore defined, Q is a leaving group such as halo, triflate; mesylate,tosylate, or alternatively, an alcohol, aldehyde or ketone, can beprepared by methods outlined in U.S. Pat. Nos. 6,002,008, 6,288,082, and6,297,258, hereby incorporated by reference. Intermediates 10, whereinr, R₆, and Het2 are herein before defined, Z is O, S or NR₆, arecommercially available, or can be prepared by methods in the chemistryliterature by a chemist skilled in the art. Alternatively, Z is a bond,r=0 Het2 possesses a nucleophilic nitrogen atom. The reaction ofcompounds of Formula 10 with compounds of Formula 9 can be carried outto provide target compounds 11 by a variety of methodologies, including:nucleophilic displacement of a leaving group, reductive amination orcoupling in the presence of reagents such as diethylazodicarboxylate andtriphenylphosphine and other methods known to chemists skilled in theart.

The preparation of the compounds and intermediates of this inventionencompassed by Formula 4 is described below in Flowsheet 5 wherein Ar,X, n, p, q, r, R₂, R₆, Z, Het1 and Het2 are herein before defined. Y isO, S or NR₆.

Compounds of Formula 12 where R₂ and X′ are hereinbefore defined, can beprepared as described in US 2001051620, hereby incorporated byreference. The reaction of7-substituted-4-oxo-1,4-dihydro-3-quinolinecarbonitriles 12 withintermediates 3, wherein p, q, r, R₆, Z, Het1 and Het2 are herein beforedefined, and Y is O, S or NR₆, provides compounds of Formula 13.Alternatively, Y is a bond, p=0 and Het1 possesses a nucleophilicnitrogen atom. Treatment of7-substituted-4-oxo-1,4-dihydro-3-quinolinecarbonitriles 13 with ahalogenating reagent PO(Z)₃ wherein Z is a chloro or bromo group whichinclude but not limited to phosphorous oxychloride, phosphorousoxybromide either neat or optionally in the presence of a cosolventwhich include but not limited to dichloromethane affords7-substituted-4-halo-3-quinolinecarbonitriles 14. Compounds of Formula14 are further reacted with a nucleophile of Formula 15 wherein Ar, nand X are as hereinbefore defined, in a solvent such as 2-ethoxyethanolin the presence of a catalytic or equivalent amount of pyridinehydrochloride, or by using bases such as trialkylamines, sodium hydridein an inert solvent, sodium or potassium alkoxides in an alcoholsolvents, and the like to provide products of Formula 1.

The preparation of the compounds and intermediates of this inventionencompassed by Formula 11 is described below in Flowsheet 6 wherein Ar,X, n, p, q, r, R₆, Z, Het1 and Het2 are herein before defined. G is R₁,R₂, R₃ or R₄. Y is O or NR₆.

Intermediates 16, wherein Ar, X and n are herein before defined, G isR₁, R₂, R₃ or R₄ and G′ is bromo, iodo or a sulfonate such as, but notrestricted to trifluoromethanesulfonate, can be prepared by methodsoutlined in U.S. Pat. Nos. 6,002,008, 6,288,082, and 6,297,258, herebyincorporated by reference. Intermediates 3, wherein p, q, r, R₆, Z, Het1and Het2 are herein before defined, and Y is O, S or NR₆, p=0 and Het1possesses a nucleophilic nitrogen atom, are commercially available, orcan be prepared by methods in the chemistry literature by a chemistskilled in the art. The reaction of compounds of Formula 3 withintermediates 16 can be carried out with an appropriately substitutedpalladium catalyst, in the presence of certain ligands (Muci, Alex R.;Buchwald, Stephen L. Practical palladium catalysts for C—N and C—O bondformation. Topics in Current Chemistry 2002, 219, 131-209) to providetarget compounds 11. Alternatively, Y is a bond, p=0 and Het1 possessesa nucleophilic nitrogen atom.

The preparation of the compounds and intermediates of this inventionencompassed by Formula 11 is described below in Flowsheet 7 wherein Ar,X, n, p, q, r, R₆, Z, Het1 and Het2 are herein before defined. G is R₁,R₂, R₃ or R₄. Y is O or NR₆.

Compounds of Formula 17 where G is R₁, R₂, R₃ or R₄ and G′ is bromo,iodo or a sulfonate such as, but not restricted totrifluoromethanesulfonate, can be prepared by methods outlined in U.S.Pat. Nos. 6,002,008, 6,288,082, and 6,297,258, hereby incorporated byreference. The reaction of7-substituted-4-oxo-1,4-dihydro-3-quinolinecarbonitriles 17 withintermediates 3, wherein p, q, r, R₆, Z, Het1 and Het2 are herein beforedefined, and Y is O or NR₆, or Y is a bond, p=0 and Het1 possesses anucleophilic nitrogen atom, provides compounds of Formula 18 whencoupled with literature methods (Muci, Alex R.; Buchwald, Stephen L.Practical palladium catalysts for C—N and C—O bond formation. Topics inCurrent Chemistry 2002, 219, 131-209). Treatment of7-substituted-4-oxo-1,4-dihydro-3-quinolinecarbonitriles 18 with ahalogenating reagent PO(Z)₃ wherein Z is a chloro or bromo group whichinclude but not limited to phosphorous oxychloride, phosphorousoxybromide either neat or optionally in the presence of a cosolventwhich include but not limited to dichloromethane affords7-substituted-4-halo-3-quinolinecarbonitriles 19. Compounds of Formula19 are further reacted with a nucleophile of Formula 15 wherein Ar, nand X are as hereinbefore defined, in a solvent such as 2-ethoxyethanolin the presence of a catalytic or equivalent amount of pyridinehydrochloride, or by using bases such as trialkylamines, sodium hydridein an inert solvent, sodium or potassium alkoxides in an alcoholsolvents, and the like to provide products of Formula 11.

In those cases where the Ar, G, R₁, R₂, R₃ and R₄ substituents maycontain an asymmetric carbon atom, the intermediates can be used as theracemate or as the individual R or S enantiomers in which case thecompounds of this invention will be in the racemic or R and S opticallyactive forms, respectively. In cases where the substituents may containmore than one asymmetric carbon atoms, diastereomers may be present;these can be separated by methods well known in the art including, butnot limited to, fractional crystallization and chromatographic methods.

Converting the R₂ groups of Formula 2, 4, 6, 12, 13 and 14 to differentR₂ groups, or converting the G groups of Formula 7, 8, 9 and 11, 16, 17,18 and 19 to various R₁, R₂, R₃ and R₄ groups can be accomplishedthrough any conventionally known techniques.

Representative compounds of this invention are evaluated in severalstandard pharmacological test procedures that showed that the compoundsof this invention possess significant activity as inhibitors ofRas/Raf/MAPK cascade and are antiproliferative agents. Disease stateswhich can be treated or inhibited by protein kinase inhibitors includethose in which the etiology is at least in part caused by a defectupstream in a signaling pathway from a protein kinase (i.e., coloncancer); those in which the etiology is at least in part caused by anoverexpressed protein kinase (i.e., lung cancer and colonic polyps); andthose in which the etiology is at least in part caused by a dysregulatedprotein kinase (gene turned on at all times; glioblastoma).

Based on the activity shown in the standard pharmacological testprocedures, the compounds of this invention are therefore useful asantineoplastic agents.

The test procedures used and results obtained are shown below.

Mitogen Activated Protein Kinase (MAPK) Test Procedure

To evaluate inhibitors of the MAP (mitogen activated protein) kinase atwo component coupled standard pharmacological test procedure, whichmeasures phosphorylation of a serine/threonine residue in an appropriatesequence in the substrate in the presence and absence of a putativeinhibitor, is used. Recombinant human MEK 1 (MAPKK) is first used toactivate recombinant human ERK2 (MAPK) and the activated MAPK (ERK) isincubated with substrate (myelin basic protein peptide (MBPP) or Mycpeptide) in the presence of ATP, Mg⁺² and radiolabeled ³³P ATP. Thephosphorylated peptide is captured on a P 81 phosphocellulose filter(paper filter or embedded in microtiter plate) ished and counted byscintillation methods.

The peptide substrates used in the assay are MBPP, peptide substrate(APRTPGGRR), or synthetic Myc substrate, (KKFELLPTPPLSPSRR•5 TFA). Therecombinant enzymes used are prepared as GST fusion proteins of humanERK 2 and human MEK 1. Inhibitor samples are prepared as 10× stocks in10% DMSO and an appropriate aliquot is used to deliver either 10 ug/mlfor a single point screening dose or 100 to 0.0001 uM finalconcentration for a dose response curve. Final DMSO concentrations areless than or equal to 1%.

The reaction is run as follows in 50 mM Tris kinase buffer, pH 7.4 in areaction volume of 50 ul. The appropriate volume of kinase buffer andinhibitor sample is added to the tube. Appropriate dilution of enzyme isdelivered to give 2-5 μg recombinant MAPK (Erk) per tube. The inhibitoris incubated with MAPK (Erk) for 30 minutes at 0° C. Recombinant Mek(MAPKK) (0.5-2.5 ug) or fully activated Mek (0.05-0.1 units) is added toactivate the Erk and incubated for 30 minutes at 30° C. Then substrateand ³³P ATP are added to give a final concentration of 0.5-1 mM MBPP or250-500 μM Myc; 0.2-0.5 μCi gamma P 33 ATP/tube; 50 μM ATP finalconcentration. Samples are incubated at 30° C. for 30 minutes and thereaction is stopped by adding 25 μl of ice cold 10% trichloroacetic acid(TCA). After samples are chilled on ice for 30 minutes, 20 μl of sampleis transferred onto P 81 phosphocellulose filter. Filter papers arewashed 2 times with a large volume of 1% acetic acid, then 2 times withwater. The filters are briefly air dried before addition of scintillantand samples are counted in the appropriate scintillation counter set upfor reading ³³P isotope. Samples included a positive control (activatedenzyme plus substrate); a no enzyme control; a no substrate control;samples with different concentrations of putative inhibitor; and sampleswith reference inhibitors (other active compounds or non-specificinhibitors such as staurosporine or K252 B).

The raw data is captured as counts per minute (cpm). Sample replicatesare averaged and corrected for background count. Mean cpm data istabulated by group and % inhibition by a test compound is calculated as(corrected cpm control−corrected. cpm sample/control)×100=% inhibition).If several concentrations of inhibitor are tested, IC₅₀ values (theconcentration which gives 50% inhibition) are determined graphically.The results obtained for representative compounds of this invention arelisted in Table 1.

Src Kinase Test Procedure

Inhibitors of p60^(c-src) (partially purified preparation purchased fromUpstate Biotechnologies) tyrosine kinase activity are analyzed in anElisa format. The Boehringer Mannheim Tyrosine Kinase Assay Kit (Catalognumber 1-534505) with a cdc2 substrate peptide containing Tyr15 is usedfor the assay. HRP-conjugated anti-phosphotyrosine is used to detectphosphorylated peptide via a color reaction. Conditions recommended bythe manufacturer are employed.

Reaction conditions: Five microliter aliquots of each compound preparedfresh at the time of the assay are added as a solution in 10 mM HEPES pH7.5, 10% DMSO to the reaction well. Thirty-five microliters of reactionmix containing Src, buffer and peptide/bovine serum albumin mix areadded to the compound wells and incubated at 30° C. for 10 minutes(reaction buffer: 50 mM Tris HCl pH 7.5, 10 mM MgCl₂, 0.1 mM EGTA, 0.5mM Na₃VO₄). The reaction is started by addition of 10 microliters ofATP, incubated at 30° C. for 1 hour, and stopped by addition of 20microliters of 0.5M EDTA. The reaction mixture with the phosphorylatedpeptide is then transferred to a streptavidin-coated microtiter plate(provided in the kit) and allowed to bind for 20 minutes. Unboundpeptide and reaction mixture is decanted and the plate is washed withPBS six times. Horseradish peroxidase-conjugated phosphotyrosineantibody supplied in the kit is incubated with the plate for one hour,then decanted. The plate is again ished with PBS six times. Substrate(provided in the kit) is added and absorbance at 405 nm is measured.

Activity is determined as % inhibition as calculated by the formula:

(1−Abs/Abs(max))×100=% inhibition. Where multiple concentrations of thetest agent are used, an IC₅₀ (concentration which gives 50% inhibition)could be determined.

The results obtained for representative compounds of this invention arelisted in Table 1

Raf/Mek Kinase Cascade Assay Procedure

Raf-1 (c-Raf) is used to phosphorylate and activate inactive GST-MEK1which then can phosphorylate and activate inactive p42 GST-MAPK, whichsubsequently is measured for phosphorylation of the TEY sequence (aa's202-204) by a phospho-specific antibody from Sigma (cat. #77439219041)Reagents: Sf9 insect cell lysate containing full length 6his-taggedrecombinant human c-Raf. (Specific Activity: ˜200 U/ml). HumanNon-active Mek-1-GST and human GST-MAP kinase (recombinant proteinsproduced in E. coli).

Stock Solutions Raf/Mek Cascade Assay:

-   1. Assay Dilution Buffer (ADB): 20 mM MOPS, pH 7.2, 25 mM β-glycerol    phosphate, 5 mM EGTA, 1 mM sodium orthovanadate, 1 mM    dithiothreitol;-   2. Magnesium/ATP Cocktail: 500 μM cold ATP and 75 mM magnesium    chloride in ADB;-   3. Active Kinase: Human Active c-Raf: Use at 0.4 U per assay point;-   4. Non-active GST-MEK1: Use at 0.1 μg per assay point; and-   5. Non-active GST-p42 MAP Kinase: Use at 1.0 μg per assay point.

Stock Solutions ELISA:

-   1. TBST-Tris (50 mM, pH 7.5), NaCl (150 mM), Tween-20 (0.05%);-   2. Superblock (Pierce);-   3. Anti-GST Ab (Pharmacia);-   4. Anti-Phospho MAPK (Sigma); and-   5. Anti-Mouse Ab/Europium conjugate (Wallac).

Assay Procedure:

First Stage: c-Raf Dependent Activation of GST-MEK and GST-MAPK

-   1. Add 20 ml of ADB per assay (i.e. per well of a 96 well plate);-   2. Add 10 ml of 0.5 mM cold ATP and 75 mM magnesium chloride in ADB;-   3. Add 2 ml of c-Raf (0.4 U/assay), in conjunction with 1.6 ml    non-active MEK1 (0.4 mg/assay);-   4. Add 4 ml of non-active GST-p42 MAP Kinase (1.0 mg/assay);-   5. Incubate for 60 minutes at 30° C. in a shaking incubator;-   6. Transfer this mixture to an anti-GST Ab coated 96 well plate    (Nunc Immunosorb plates coated o/n with a-GST, then blocked with    Pierce Superblock);-   7. Incubate for 60 minutes at 30° C. in a shaking incubator;-   8. Wash 3× with TBST, add Anti-Phospho MAPK (Sigma) (1:3000);-   9. Incubate for 60 minutes at 30° C. in a shaking incubator;-   10. Wash 3× with TBST, add Anti-Mouse Ab/Europium conjugate (Wallac)    (1:500);-   11. Incubate for 60 minutes at 30° C. in a shaking incubator;-   12. Wash 3× with TBST, Read plates in Wallac Victor model Plate    Reader; and-   13. Collect data analyze in Excel for single point and IC₅₀    determinations.

Single point assay—% inhibition at 10 mg/ml (% Inhibition=1−cpd. treatedsample/untreated control). IC₅₀ determinations—done on compounds fromsingle point assays with >80% inhibition. Typically, the Raf/Mek assayis run at compound concentrations from 10 μM to 1 nM in half logdilutions. (% inhibition is determined for each compound concentration).The results obtained measure Raf and/or Mek kinase inhibition, and forrepresentative compounds of this invention are listed in Table 1.

TABLE 1 Inhibition of Raf and/or Mek kinase (Raf/Mek), Mitogen ActivatedProtein Kinase (Mek-Erk) and p60^(c-src) (Src) Raf/Mek Mek-Erk SrcExample IC50 (μM) IC50 (μM) IC50 (μM) 11 0.114 (b) 0.281 (a) 12 0.210(c) 13 1.427 14 0.795 (a) 15 1.721 16 0.035 17 0.320 18 0.075 19 0.19920 0.293 21 0.181 22 0.273 23 0.188 28 2.390 29 1.644 (b) 30 0.220 (a)32 0.020 0.045 33 0.003 34 0.250 0.00055 35 0.001 (a) Average of tworuns (b) Average of three runs (c) Average of four runs

Cell Proliferation Test Procedure for Inhibitors of Src Kinase

HT-29 cells: Compound effectiveness at inhibiting cell proliferation onplastic is performed in a 96-well format by plating 5000 cells per wellin appropriate medium on day one, followed by compound addition on day 2in serial two-fold dilutions. On day five, compound is washed away andmedium containing MTS reagent (Promega) is added. Relative cell numberis determined by reading the absorbance at 490 nm of a dye produced byan NAD-dependent cellular enzymatic reaction. These data are shown belowin Table 2.

Anchorage Independent Src-transformed Fibroblast Proliferation TestProcedure: Rat2 fibroblasts stably transformed with a plasmid containinga CMV promotor controlled v-Src/Hu c-Src fusion gene in which thecatalytic domain of human c-Src is inserted in place of the v-Srccatalytic domain in the v-Src gene are used for the measurement of srcdependent suspension growth. Ultra-low cluster plates (Costar #3474) areseeded with 10,000 cells per well on Day 1. Compound is added in serialtwo-fold dilutions from 10 micromolar to 0.009 micromolar on Day 2 andMTS reagent (Promega) is added on Day 5 (100 microliters of MTS/mediummix+100 microliters of medium already on the cells and the absorbance ismeasured at 490 nm. The results are analyzed as follows to yield an IC₅₀for proliferation (micromolar units) as follows: % inhibition=(Abs490 nmsample−blank)/(Abs490 nm no cmpd control−blank)×100%. These data areshown below in Table 2.

TABLE 2 Inhibition of Cancer Cell Growth HT-29 prolif Src TF prolifExample IC₅₀ (μM) IC₅₀ (μM) 31 2.4 34 2.6 0.466 35 2.7 0.140Cell Based Screen for Inhibitors of Raf and/or Mek Kinase.

Materials

Cell Lines: Human adenocarcinoma cell line LoVo, known to be growthinhibited by low nM concentrations of a reference standard inhibitor ofRas and human adenocarcinoma cell line CaCo-2, known to be growthresistant to the same reference compound.

Cell Media: RPMI 1640 with 10% Fetal Bovine Serum supplemented withL-glutamine and Penicillin/Streptomycin.

Compounds: Supplied usually as a 10 mM stock in 100% DMSO.

Normal Saline: 150 mM NaCl

Trichloroacetic Acid (TCA): 50% (w/v) in water

Sulforhodamine B (SRB): 0.4% (w/v) in 1% Acetic Acid

Tris Base: 10 mM in water

Methods

Cells are plated at 2000 cells per well for cell line LoVo and 1500cells for cell line CaCo-2 in 96 well plates. Cells are plated in media(200 μl) and allowed to adhere overnight at 37° C. At 24 hours postplating, compounds are added directly at a volume of 0.5 μl. For thequalitative screen (compounds screened at 25 μM) compound is addeddirectly to cells. For the quantitative screen, compound is firstdiluted in DMSO to generate concentrations of compound or referencestandard of: 1, 5, 10 and 25 μM. It is advisable to make the dilutionsin an identical 96 well plate so that compounds can be added using amultichannel micropipettor set at 0.5 μl. The cells are then incubatedfor four days after which the media is removed using a 12 well manifoldby first tipping the plate forward at a 45 degree angle and theninserting the manifold in an upright orientation to prevent the tips ofthe manifold from disturbing cells at the bottom of the plate. 200 μl ofnormal saline is then added to each well using an 8 well multichannelpipettor, followed by the careful addition of 50 μl of 50% TCA. Theplates are then incubated for 2 hours at 4° C., after which thesupernatant is removed using the same technique as above and the platedcells are washed twice with 200 μl water. The plates are then air driedand 50 μl of SRB stock solution is carefully added so that the entirebottom of each well is covered. This again can be done using an 8 wellmultichannel pipettor. The SRB is incubated with fixed cells for 15minutes at room temperature after which the SRB is removed with themanifold as described above and the plates washed twice with 350 μl of1% acetic acid per well each time. The plates are then air dried afterwhich the bound SRB is released from protein by the addition of 200 μlof Tris base. Resolubilizing the SRB is aided by placing the plates on arotator for 15-30 minutes. The absorbance of each well is determined at550 or 562 nm using a microtiter plate reader.

Each compound or dilution thereof is performed in triplicate. Outliersare identified by visual inspection of the data. Each plate should havea “0” control (vehicle only).

Qualitative screen: To calculate % inhibition of a compound at 25 μM,the following formula is used: 1−(experimental absorbance @ 25 μMcompound/“0” control absorbance)×100=% inhibition at 25 μM. Compoundshaving >50% inhibition at 25 μM are placed in the quantitative assay.

Quantitative Assay: A standard curve is constructed by plotting theconcentration of compound against the average absorbance calculated atthat concentration. A curve is plotted and the concentration at whichthe curve passes through the 50% the absorbance mark seen in the “0”control well is the IC₅₀ calculated for that compound. Multiple entriesfor a given compound indicate that it is tested multiple times. Theresults obtained for representative compounds of this invention arelisted in Table 3.

TABLE 3 LoVo BxPC3 Example IC50 μM IC50 μM 11 0.029 (c) 0.042 (b) 120.46 (b) 1.4 13 0.47 0.6 14 0.37 0.3 15 0.261 0.36 16 0.009 0.03 (b) 170.031 18 0.027 0.03 19 0.008 20 0.022 0.038 21 0.02 22 0.08 23 0.474 280.014 0.008 (b) 29 0.004 0.008 (a) 30 0.014 0.03 32 0.9 33 0.047 0.049(a) (a) Average of two runs (b) Average of three runs (c) Average offive runs

The results shown in tables 1, 2, 3, 4 and 5 demonstrate that thecompounds of this invention are potent inhibitors of protein kinases,and are useful as described above.

The preparation of representative examples of the compounds of thisinvention is described below.

EXAMPLE 14-[3-Chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrile

A mixture of 1.0 g (4.23 mmol) of4-chloro-7-fluoro-6-methoxyquinoline-3-carbonitrile (AM100856), 1.114 g(4.65 mmol) of3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamine and 0.489 g(4.23 mmol) of pyridine hydrochloride in 10 mL of 2-ethoxyethanol isheated at 120° C. for 1.5 hours, then cooled to room temperature. Theresulting solid is filtered, washed with cold 2-ethoxyethanol, thenethyl acetate. After drying in vacuo, the solid is suspended in asaturated solution of sodium bicarbonate, stirred for 45 minutes andcollected by filtration. The reaction product is washed with water anddried in vacuo, to provide4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileas a tan solid, mp 295-297° C.

MS (ES, positive ion mode): m/z calcd for C₂₁H₁₅ClFN₅OS: 439.1, found:439.8 (M+H)⁺.

EXAMPLE 27-Fluoro-6-methoxy-4-[4-(pyridin-2-ylsulfanyl)-phenylamino]quinoline-3-carbonitrile

Following the procedure of Example 1, a mixture of 635.4 mg (2.68 mmol)4-chloro-7-fluoro-6-methoxy-quinoline-3-carbonitrile, 542.1 mg (2.81mmol) of 4-(pyridin-2-ylsulfanyl)phenylamine and 309.8 mg (2.68 mmol) ofpyridine hydrochloride are refluxed in 2-ethoxyethanol (27 mL) at 100°C. for 12 hours to provide 830 mg of7-fluoro-6-methoxy-4-[4-(pyridin-2-ylsulfanyl)-phenylamino]quinoline-3-carbonitrileas an off-white solid, mp 231-233° C.

MS (ES, positive mode): m/z calcd for C₂₂H₁₅FN₄OS: 402.1, found 403.2(M+1).

EXAMPLE 34-[3-Chloro-4-pyridin-2-ylsulfanyl)phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrile

Following the procedure of Example 1, a mixture of 635.4 mg (2.68 mmol)4-chloro-7-fluoro-6-methoxy-quinoline-3-carbonitrile, 642.1 mg (2.81mmol) of 3-chloro-4-(pyridin-2-ylsulfanyl)-phenylamine and 309.8 mg(2.68 mmol) of pyridine hydrochloride are refluxed in 2-ethoxyethanol(27 mL) at 100° C. for 12 hours to yield4-[3-chloro-4-(pyridin-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileas a yellow solid, mp 225-228° C.

MS (ES, positive mode): m/z calcd for C₂₂H₁₄ClFN₄OS: 436.1, found 437.0(M+1).

EXAMPLE 47-Fluoro-6-methoxy-4-[4-pyridin-3-yloxy)-phenylamino]-quinoline-carbonitrile

Following the procedure of Example 1, a mixture of 635.4 mg (2.68 mmol)4-chloro-7-fluoro-6-methoxy-quinoline-3-carbonitrile, 500 mg (2.81 mmol)of 4-(pyridin-3-yloxy)phenylamine and 309.8 mg (2.68 mmol) of pyridinehydrochloride are refluxed in 2-ethoxyethanol (27 mL) at 100° C. for 12hours to yield7-fluoro-6-methoxy-4-[4-(pyridin-3-yloxy)-phenylamino]-quinoline-3-carbonitrileas an off-white solid, mp 235-236° C.

MS (ES, positive mode): m/z calcd for C₂₂H₁₅FN₄O₂: 386.1, found 387.2(M+1).

EXAMPLE 57-Fluoro-6-methoxy-4-[(2-methoxy-phenoxy)phenylamino]quinoline-3-carbonitrile

Following the procedure of Example 1, a mixture of 626 mg (2.65 mmol)4-chloro-7-fluoro-6-methoxy-quinoline-3-carbonitrile, 600 mg (2.79 mmol)of7-fluoro-6-methoxy-4-[4-(2-methoxyphenoxy)phenylamino]quinoline-3-carbonitrileand 310 mg (2.65 mmol) of pyridine hydrochloride are refluxed in2-ethoxyethanol (7 mL) at 120° C. for 30 minutes to yield7-fluoro-6-methoxy-4-[4-(2-methoxy-phenoxy)phenylamino]quinoline-3-carbonitrileas a white solid, mp 146-150° C.

MS (ES, positive mode): m/z calcd for C₂₄H₁₈FN₃O₃: 415.4, found 416.2(M+1).

EXAMPLE 64-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-fluoroquinoline-3-carbonitrile

Following the procedure of Example 1, a mixture of 1.2 g (5.81 mmol)4-chloro-7-fluoro-3-quinolinecarbonitrile, 1.53 g (6.39 mmol) of3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamine and 0.671 g(5.81 mmol) of pyridine hydrochloride are refluxed in 2-ethoxyethanol(12 mL) at 120° C. for 45 minutes to yield4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-fluoroquinoline-3-carbonitrileas an off-white solid, mp 268-272° C.

MS (ES, negative mode): m/z calcd for C₂₀H₁₃ClFN₅S: 409.9, found 408.1,410.1 (M−1).

EXAMPLE 74-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]phenyl}amino)-6-ethoxy-7-fluoro-3-quinolinecarbonitrile

Following the procedure of Example 1, a mixture of 2.4 g (9.72 mmol)4-chloro-4-ethoxy-7-fluoro-3-quinolinecarbonitrile, 2.56 g (10.7 mmol)of 3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamine and 1.2 g(10.38 mmol) of pyridine hydrochloride are refluxed in 2-ethoxyethanol(30 mL) at 110° C. for 1 hour to yield4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]phenyl}amino)-6-ethoxy-7-fluoro-3-quinolinecarbonitrileas a pink solid, mp 251-254° C.

MS (ES, positive mode): m/z calcd for C₂₂H₁₇ClFN₅OS: 453.9, found 454.3,456.3 (M+1).

EXAMPLE 87-(2-Chloroethoxy)-6-methoxy-4-(4-phenoxyphenylamino)quinoline-3-carbonitrile

Following the procedure of Example 1, a mixture of 3.0 g (0.01 mol) of4-chloro-7-(2-chloroethoxy)-6-methoxy-3-quinolinecarbonitrile(Boschelli, Diane H.; Ye, Fei; Wang, Yanong D.; Dutia, Minu; Johnson,Steve L.; Wu, Biqi; Miller, Karen; Powell, Dennis W.; Yaczko, Deanna;Young, Mairead; Tischler, Mark; Arndt, Kim; Discafani, Carolyn; Etienne,Carlo; Gibbons, Jay; Grod, Janet; Lucas, Judy; Weber, Jennifer M.;Boschelli, Frank. J. Med. Chem. 2001, 44, 3965-3977), 2.04 g (0.011 mol)of 4-phenoxyaniline and pyridine hydrochloride 1.16 g (0.01 mol) in 100mL of 2-ethoxyethanol is heated at 135° C. for 2 hours. Water is addedto the reaction mixture, and the precipitate is filtered off. Afterwashing with water, ether and ethyl acetate, the solid is dried in vacuoto provide 3.48 g of7-(2-chloroethoxy)-6-methoxy-4-(4-phenoxyphenylamino)quinoline-3-carbonitrileas a yellow solid, mp 148-151° C.

MS (ES, positive ion mode): m/z calcd for C₂₆H₂₀ClN₃O₃: 445.12, found:446.0, 448.0 (M+1).

EXAMPLE 94-{3-Chloro-4-[1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-7-(3-chloropropoxy)-6-methoxy-3-quinolinecarbonitrile

A 2.0 g (6.43 mmol) portion of4-chloro-7-(3-chloropropoxy-6-methoxy-3-quinolinecarbonitrile(Boschelli, Diane H.; Ye, Fei; Wang, Yanong D.; Dutia, Minu; Johnson,Steve L.; Wu, Biqi; Miller, Karen; Powell, Dennis W.; Yaczko, Deanna;Young, Mairead; Tischler, Mark; Arndt, Kim; Discafani, Carolyn; Etienne,Carlo; Gibbons, Jay; Grod, Janet; Lucas, Judy; Weber, Jennifer M.;Boschelli, Frank. J. Med. Chem. 2001, 44, 3965-3977) in 2-ethoxyethanol(20 mL) is heated to 60° C. To this is added 1.7 g (7.10 mmol) of3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamine (U.S. Pat.No. 4,973,599) and 0.74 g (6.43 mmol) of pyridine hydrochloride, and theresulting mixture is refluxed for 2 hours. After cooling, theprecipitated solid is filtered, washed with aqueous sodium bicarbonateand dried in vacuo to give 2.93 g of4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-7-(3-chloropropoxy)-6-methoxy-3-quinolinecarbonitrileas a tan solid, mp 271-276° C.

MS (ES, positive ion mode): m/z calcd for C₂₄H₂₁Cl₂N₅O₂S: 514.5, found:514.1, 516.1 (M+1).

EXAMPLE 107-(2-Chloroethoxy)-4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-3-quinolinecarbonitrile

Following the procedure of Example 9, 1.0 g (3.36 mmol) of4-chloro-7-(2-chloroethoxy)-6-methoxy-3-quinolinecarbonitrile(Boschelli, Diane H.; Ye, Fei; Wang, Yanong D.; Dutia, Minu; Johnson,Steve L.; Wu, Biqi; Miller, Karen; Powell, Dennis W.; Yaczko, Deanna;Young, Mairead; Tischler, Mark; Arndt, Kim; Discafani, Carolyn; Etienne,Carlo; Gibbons, Jay; Grod, Janet; Lucas, Judy; Weber, Jennifer M.;Boschelli, Frank. J. Med. Chem. 2001, 44, 3965-3977) is reacted with 0.9g (3.7 mmol) of3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamine (U.S. Pat.No. 4,973,599) and 0.4 g (3.46 mmol) of pyridine hydrochloride in2-ethoxyethanol (10 mL) to provide 1.44 g of7-(2-chloroethoxy)-4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-3-quinolinecarbonitrileas a tan solid, mp 278-280° C.

MS (ES, positive ion mode): m/z calcd for C₂₃H₁₉Cl₂N₅O₂S: 500.4, found:499.8, 501.7 (M+1).

EXAMPLE 114-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile

A mixture of 150 mg (0.34 mmol) of4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileand 316 mg (2.05 mmol) of 4-pyrrolidin-1-yl-piperidine in 1 mL of1-methyl-2-pyrrolidinone is heated at 105° C. for 16 hours. The solventsare removed in vacuo. A 10 mL portion of water is added to the residue,from which a tan solid is precipitated out. The solid is filtered offand washed with water. After drying in vacuo, the solid is suspended inethyl acetate and stirred for 1 hour. The solid is filtered off, washedwith ethyl acetate and dried in vacuo to provide 140 mg of4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-6-methoxy-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrileas a yellow solid, mp 225-229° C.

MS (ES, positive ion mode): m/z calcd for C₃₀H₃₂ClN₇OS: 573.2, found:574.1, 576.1 (M+1).

EXAMPLE 126-Methoxy-4-{[4-(2-methoxyphenoxy)phenyl]amino}-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile,trifluoroacetate salt

Following the procedure of Example 11, a mixture of 50 mg (0.12 mmol) of7-fluoro-6-methoxy-4-[4-(2-methoxy-phenoxy)phenylamino]quinoline-3-carbonitrileand 111 mg (0.72 mmol) of 4-(1-pyrrolidinylpiperidine) in 0.5 mL of1-methyl 2-pyrrolidinone is heated at 105° C. for 28 hours to yield thecrude product. Purification by a Gilson HPLC (gradient solvents) gives20 mg of6-methoxy-4-{[4-(2-methoxyphenoxy)phenyl]amino}-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile,trifluoroacetate salt as a brown oil.

MS (ES, positive ion mode): m/z calcd for C₃₃H₃₅N₅O₃: 549.7, found:550.5 (M+1).

EXAMPLE 136-Methoxy-4-[4-(pyridin-2-ylsulfanyl)phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)-quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 150 mg (0.37 mmol)of7-fluoro-6-methoxy-4-[4-pyridin-2-ylsulfanyl)-phenylamino]-quinoline-3-carbonitrile,287 mg (1.86 mmol) of 4-(1-pyrrolidinylpiperidine) are refluxed in1-methyl 2-pyrrolidinone (1 mL) at 105° C. for 12 hours to yield thecrude product. Purification by silica gel chromatography (95:5 methylenechloride/methanol) gives 107 mg of6-methoxy-4-[4-(pyridin-2-ylsulfanyl)-phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrileas a yellow solid, mp 182-184° C.

MS (ES, positive mode): m/z calcd for C₃₁H₃₂N₆OS: 536.2, found 537.3(M+1).

EXAMPLE 144-[3-Chloro-4-(pyridin-2-ylsulfanyl)-phenylamino]-6-methoxy-7-(4-pyrrolidin-1-yl-piperidin-1-yl)-quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 150 mg (0.34 mmol)of4-[3-Chloro-4-(pyridin-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileand 264 mg (1.71 mmol) of 4-(1-pyrrolidinylpiperidine) are refluxed in1-methyl 2-pyrrolidinone (1 mL) at 105° C. for 12 hours to yield thecrude product. Purification by silica gel chromatography (95:5 methylenechloride/methanol) gives 126 mg of4-[3-chloro-4-(pyridin-2-ylsulfanyl)-phenylamino]-6-methoxy-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrileas a yellow solid, mp 203-205° C.

MS (ES, positive mode): m/z calcd for C₃₁H₃₁ClN₆OS: 570.2, found 571.3(M+1).

EXAMPLE 156-Methoxy-4-[4-(pyridin-3-yloxy)-phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)-quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 150 mg (0.39 mmol)of7-fluoro-6-methoxy-4-[4-(pyridin-3-yloxy)phenylamino]-quinoline-3-carbonitrileand 299 my (1.94 mmol) of 4-(1-pyrrolidinylpiperidine) are refluxed in1-methyl 2-pyrrolidinone (1 mL) at 105° C. for 12 hours to yield thecrude product. Purification by silica gel chromatography (95:5 methylenechloride/methanol) gives 120 my of6-methoxy-4-[4-(pyridin-3-yloxy)phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-ylquinoline-3-carbonitrileas a yellow solid, mp 194-196° C.

MS (ES, positive mode): m/z calcd for C₃₁H₃₂N₆O₂: 520.6, found 521.3(M+1).

EXAMPLE 164-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-(4-pyrrolidin-1-ylpiperidin-1-ylquinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 150 mg (0.37 mmol)of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-fluoroquinoline-3-carbonitrileand 339 mg (2.20 mmol) of 4-(1-pyrrolidinylpiperidine) in 1 mL of1-methyl 2-pyrrolidinone is heated at 105° C. for 17 hours to yield thecrude product. Purification by silica gel chromatography (gradient 98:2methylene chloride/methanol to 9:1 methylene chloride/methanol) gives 66mg of4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrileas a yellow solid, mp 125-130° C.

MS (ES, negative ion mode): m/z calcd for C₂₉H₃₀ClN₇S: 544.1, found:542.1, 543.9 (M−1).

EXAMPLE 174-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-(2-methoxyethoxy)-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 150 mg (0.31 mmol)of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]phenyl}amino)-7-fluoro-6-(2-methoxyethoxy)-3-quinolinecarbonitrileand 287 mg (1.86 mmol) of 4-(1-pyrrolidinylpiperidine) in 1 mL of1-methyl 2-pyrrolidinone is heated at 105° C. for 17 hours to yield thecrude product. Purification by silica gel chromatography (gradient 98:2methylene chloride/methanol to 4:1 methylene chloride/methanol) gives120 mg of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-(2-methoxyethoxy)-7-(4-pyrrolidin-1-ylpiperidin-1-ylquinoline-3-carbonitrile as a yellow solid, mp 238-241° C.

MS (ES, positive ion mode): m/z calcd for C₃₂H₃₆ClN₇O₂S: 635.2, found:635.4, 637.3 (M+1).

EXAMPLE 184-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-ethoxy-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 150 mg (0.33 mmol)of 4-({3chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]phenyl}amino)-6-ethoxy-7-fluoro-3-quinolinecarbonitrileand 306 mg (1.98 mmol) of 4-(1-pyrrolidinylpiperidine) in 1 mL of1-methyl 2-pyrrolidinone is heated at 105° C. for 17 hours to yield thecrude product. Purification by silica gel chromatography (gradient 98:2methylene chloride/methanol to 4:1 methylene chloride/methanol) gives122 mg of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-ethoxy-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrileas a yellow solid, mp 203-207° C.

MS (ES, positive ion mode): m/z calcd for C₃₁H₃₄ClN₇OS: 588.2, found:588.3, 590.4 (M+1).

EXAMPLE 197-(1,4′-Bipiperidin-1′-yl)-4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-4-methoxyquinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 220 mg (0.5 mmol) of4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileand 290 mg (1.72 mmol) of 4-piperidinopiperidine in 1.5 mL of 1-methyl2-pyrrolidinone is heated at 105° C. for 17 hours to yield the crudeproduct. Purification by silica gel chromatography (gradient 94:6methylene chloride/methanol to 89:10:1 methylenechloride/methanol/ammonium hydroxide) gives 193 mg of7-(1,4′-bipiperidin-1′-yl)-4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxyquinoline-3-carbonitrileas a beige solid, mp 221-223° C.

MS (ES, positive ion mode): m/z calcd for C₃₁H₃₄ClN₇OS: 588.2, found:588.6 (M+1).

EXAMPLE 204-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 110 mg (0.25 mmol)of4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileand 140 mg (0.76 mmol) of 1-methyl-4-piperidin-4-ylpiperazine in 0.8 mLof 1-methyl 2-pyrrolidinone is heated at 105° C. for 28 hours to yieldthe crude product. Purification by silica gel chromatography (gradient94:6 methylene chloride/methanol to 84:15:1 methylenechloride/methanol/ammonium hydroxide) gives 100 mg of the4-({3-chloro-4-[(1-methyl-1H-imidazole-2-ylthio]phenyl}amino)-6-methoxy-7-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]quinoline-3-carbonitrileas a beige solid, mp 235-237° C.

MS (ES, positive ion mode): m/z calcd for C₃₁H₃₅ClN₈OS: 603.2, found:603.2 (M+1).

EXAMPLE 214-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-(4-morpholin-4-ylpiperidin-1-yl)quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 150 mg (0.33 mmol)of 4-[3chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileand 170 mg (1.0 mmol) of 1-methyl-4-piperidin-4-ylmorpholine in 1.0 mLof 1-methyl 2-pyrrolidinone is heated at 105° C. for 24 hours to yieldthe crude product. Purification by silica gel chromatography (gradient97:3 methylene chloride/methanol to 9:1 methylene chloride/methanol)gave 100 mg of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-(4-morpholin-4-ylpiperidin-1-yl)quinoline-3-carbonitrileas a beige solid, mp 219-221° C.

MS (ES, positive ion mode): m/z calcd for C₃₀H₃₂ClN₇O₂S: 590.2, found:590.2, 592.1 (M+1).

EXAMPLE 224-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-[4-(1-methylpiperidin-4-yl)piperazin-1-yl]quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 220 mg (0.5 mmol) of4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrile,290 mg (1.6 mmol) of 1-(1-methylpiperidin-4-yl)piperazine in 1.2 mL of1-methyl 2-pyrrolidinone is heated at 105° C. for 20 hours to yield thecrude product. Purification by silica gel chromatography (gradient 9:1methylene chloride/methanol to 85:15 methylene chloride/methanol) gives210 mg of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-methoxy-7-[4-(1-methylpiperidin-4-yl)piperazin-1-yl]quinoline-3-carbonitrile as a beige solid,mp 220-226° C. (shrinks at 205° C.).

MS (ES, positive ion mode): m/z calcd for C₃₁H₃₅ClN₈OS: 603.2, found:603.2 (M+1).

EXAMPLE 234-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-{4-[2-(1H-imidazole-1-yl)ethyl]piperazin-1-yl}-6-methoxyquinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 220 mg (0.5 mmol) of4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-7-fluoro-6-methoxyquinoline-3-carbonitrileand 290 mg (1.6 mmol) of 1-(2-imidazole-1-yl-ethyl)piperazine in 1.2 mLof 1-methyl 2-pyrrolidinone is heated at 105° C. for 20 hours to yieldthe crude product. Purification by silica gel chromatography (gradient95:5-methylene chloride/methanol to 85:15 methylene chloride/methanol)gives 210 mg of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-7-{4-[2-1H-imidazole-1-yl)ethyl]piperazin-1-yl}-6-methoxyquinoline-3-carbonitrileas a beige solid, mp 251-253° C.

MS (ES, positive ion mode): m/z calcd for C₃₀H₃₀ClN₉OS: 600.1, found:600.2 (M+1).

EXAMPLE 246-Nitro-4-oxo-7-(4-pyrrolidin-1-ylpiperidin-1-yl)-1,4-dihydroquinoline-3-carbonitrile

A mixture of 1.3 g (5.21 mmol) of7-chloro-4-hydroxy-6-nitro-3-quinolinecarbonitrile (U.S. Pat. No.6,297,258) and 2.6 g (16.67 mmol) of 4-(1-pyrrolidine)piperidine 17.0 mlof 1-methyl 2-pyrrolidinone is heated at 120° C. for 5 hours. Themixture is cooled to room temperature and stirred with 15 mL ofsaturated solution of sodium bicarbonate. Following removal of solventsin vacuo, a residue is obtained, which is stirred with ether. The solidis collected by filtration and washed with ether. The crude product isdissolved in 9:1 methylene chloride/methanol, filtered through a pad ofCelite (to remove inorganic material) and further washed with 9:1methylene chloride/methanol. Evaporation of the filtrate to drynessyields a glassy residue. Purification by silica gel chromatography(gradient 9:1 methylene chloride/methanol to 4:1 methylenechloride/methanol) gives 1.3 g of6-nitro-4-oxo-7-(4-pyrrolidin-1-ylpiperidin-1-yl)-1,4-dihydroquinoline-3-carbonitrileas an orange solid, mp 232-235° C.

MS (ES, positive ion mode): m/z calcd for C₁₉H₂₁N₅O₃: 367.4, found:368.2 (M+1).

EXAMPLE 254-chloro-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile

A mixture of 1.3 g (3.54 mmol) of6-nitr2o-4-oxo-7-(4-pyrrolidin-1-ylpiperidin-1-yl)-1,4-dihydroquinoline-3-carbonitrile,20 ml of oxalyl chloride and 10 drops of dimethyl formamide in 20 ml ofmethylene chloride is heated under reflux for 2 hours. After allowingthe mixture to cool to room temperature, the solvent is evaporated todryness in vacuo. Toluene is added to the resulting residue andevaporated in vacuo. Following a second toluene addition/evaporationcycle, the resulting solid is cooled in an ice bath and neutralized withsaturated solution of sodium bicarbonate. The resulting solid iscollected by filtration and dried in vacuo to give 1.25 g of4-chloro-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrileas a yellow solid, mp 255-265 (dec).

MS (ES, positive ion mode): m/z calcd for C19H20ClN5O2: 385.9, found:386.2 (M+1).

EXAMPLE 264-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile

A mixture of 0.5 g (1.45 mmol) of4-chloro-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile,0.38 g (1.6 mmol) of3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]aniline and 0.17 g ofpyridine hydrochloride in 7.0 mL of 2-ethoxyethanol is heated at 105° C.for 1 hour. After cooling to room temperature, the mixture is stirredwith 15 mL of saturated solution of sodium bicarbonate. The resultingsolid is collected by filtration and washed with water. Some of theproduct dissolves in water, which is recovered by extracting the aqueouslayer with solution of 95:5 methylene chloride/methanol. The organiclayer is evaporated to provide crude product. The combined solids arepurified by silica gel chromatography (gradient 95:5 methylenechloride/methanol to 4:1 methylene chloride/methanol) gives 0.52 g of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-ylquinoline-3-carbonitrileas an orange solid, mp 220-230° C.

MS (ES, positive ion mode): m/z calcd for C₂₉H₂₉ClN₈O₂S: 589.1, found:589.2 (M+H).

EXAMPLE 276-Amino-4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrile

To a refluxing mixture of 0.5 g (8.95 mmol) of iron powder, 0.7 g (13.1mmol) of ammonium chloride and 14 mL of water is added 1.26 g (2.14mmol) of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-nitro-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrilein portions so as to maintain gentle reflux. The resulting mixture isheated under reflux for 45 minutes. The mixture is filtered hot througha pad of Celite, washed with ethanol, and the filtrate is evaporated todryness to yield a residue. A saturated solution of sodium bicarbonateis added to the residue, and the mixture is stirred. The solid iscollected by filtration, washed with water and dried to yield 1.2 g of6-amino-4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrileas a yellow solid, mp 265-267° C.

MS (ES, positive ion mode): m/z calcd for C₂₉H₃₁ClN₈S: 559.1, found:559.2 (M+1).

EXAMPLE 28N-Acetyl-N-[4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-3-cyano-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinolin-6-yl]acetamide

To a cold (0° C.-5° C.) solution of 1.2 g (2.14 mmol) of6-amino-4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrileand 3.6 ml (22.6 mmol) of N,N-diethyl aniline in 18 mL of 1-methyl2-pyrrolidinone is added dropwise 1.6 ml (21.2 mmol) of acetyl chloride.The resulting mixture is stirred at room temperature for 4 hours. Thesolvent is removed in vacuo to yield a residue, which is triturated withether. A saturated solution of sodium bicarbonate is added to theresulting solid and the mixture is stirred, collected by filtration anddried in vacuo. Purification by silica gel chromatography (gradient 95:5methylene chloride/methanol to 4:1 methylene chloride/methanol) gives738 mg ofN-acetyl-N-[4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)-phenylamino]-3-cyano-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinolin-6-yl]acetamideas a beige solid, mp 245-247° C.

MS (ES, positive ion mode): m/z calcd for C₃₃H₃₅ClN₈O₂S: 643.2, found:643.2 (M+1).

EXAMPLE 29N-[4-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-3-cyano-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinolin-6-yl]acetamide

By the procedure of Example 27, 0.1 g (0.18 mmol) of4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-amino-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile is reacted with 0.3 ml (1.9mmol) of N,N-diethyl aniline and 0.04 ml (0.56 mmol) of acetyl chloridein 1.5 mL of NMP. The crude residue is purified by silica gelchromatography (gradient 95:5 methylene chloride/methanol to 4:1methylene chloride/methanol) to give 95 mg ofN-[4-({3-chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-3-cyano-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinolin-6-yl]acetamideas a beige solid, mp 243-255° C.

MS (ES, positive ion mode): m/z calcd for C₃₁H₃₃ClN₈OS: 601.2, found:601.5 (M+1).

EXAMPLE 304-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-(methylamino)-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile

A mixture of 130 mg (0.23 mmol) of6-amino-4-[3-chloro-4-(1-methyl-1H-imidazole-2-ylsulfanyl)phenylamino]-7-(4-pyrrolidin-1-yl-piperidin-1-yl)quinoline-3-carbonitrile,12.0 mg (0.4 mmol) of paraformaldehyde, 25.0 mg (0.39 mmol) of sodiumcyanoborohydride and 2 drops of acetic acid in 15 mL of ethanol isheated under reflux for 8 hours. The mixture is cooled to roomtemperature and solvent removed in vacuo. A 12 mL solution of 1N sodiumhydroxide is added to the residue, and the mixture is stirred. A crudesolid is collected by filtration, washed with water and dried.Purification by silica gel chromatography (gradient 95:5 methylenechloride/methanol to 4:1 methylene chloride/methanol) gives 66.0 mg ofthe4-({3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)thio]phenyl}amino)-6-(methylamino)-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrileas an yellow solid, mp 242-250° C.

MS (ES, positive ion mode): m/z calcd for C₃₀H₃₃ClN₈S: 573.2, found:573.3 (M+1).

EXAMPLE 314-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-(4-pyrrolidin-1-ylpiperidin-1-yl)quinoline-3-carbonitrile

Following the procedure of Example 11, a mixture of 200.0 mg (0.51 mmol)of4-(2,4-dichloro-5-methoxyanilino)-7-fluoro-6-methoxy-3-quinolinecarbonitrileand 472.0 mg (3.06 mmol) of 4-(1-pyrrolidinyl)piperidine in 1.0 mL of1-methyl-2-pyrrolidinone is heated in a sealed tube at 105° C. for 15hours to yield the crude product. Purification by preparative thin layerchromatography (eluting with 4:1 dichloromethane/methanol) provides116.5 mg of the pure product as a yellow solid, mp 225-227° C.

MS (ES, negative ion mode): m/z calcd for C₂₇H₂₉Cl₂N₅O₂: 525.2, found:524.3 (M−1).

EXAMPLE 327-(2-[1,4′]Bipiperidinyl-1′-yl-ethoxy)-6-methoxy-4-(4-phenoxy-phenylamino)-quinoline-3-carbonitrile

A mixture of 222.5 mg (0.5 mmol) of7-(2-chloroethoxy)-6-methoxy-4-(4-phenoxyphenylamino)quinolin-3-carbonitrile,841.4 mg (5.0 mmol) of 4-piperidinopiperidine and 49.5 mg (0.33 mmol) ofsodium iodide in 6.0 mL of ethylene glycol dimethyl ether are heated ina sealed tube at 135-140° C. for 17 hours to yield the crude product.Solvents are removed in vacuo. Purification of the oily residue iscarried out by preparative thin layer chromatography (eluting with 85:15dichloromethane/methanol) to provide 137.7 mg of742-[1,4′]bipiperidinyl-1′-yl-ethoxy)-6-methoxy-4-(4-phenoxy-phenylamino)-quinoline-3-carbonitrileas a light yellow solid, mp 141° C. (decompose).

MS (ES, positive ion mode): m/z calcd for C₃₅H₃₉N₅O₃: 577.3, found:578.1 (M+1).

EXAMPLE 334-{3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrile

A mixture of 150.0 mg (0.29 mmol) of4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-7-(3-chloropropoxy)-6-methoxy-3-quinolinecarbonitrileand 43.7 mg (0.29 mmol) of sodium iodide in 4.0 mL of piperidine areheated in a sealed tube at 115° C. for 5 hours to yield the crudeproduct. After removal of the solvents in vacuo, water is added to theresidue to precipitate out the crude product. The solid is filtered,then dissolved in dichloromethane/methanol. Purification is carried outby preparative thin layer chromatography (eluting with 9:1:0.1dichloromethane/methanol/aqueous ammonium hydroxide) to provide 56.8 mgof4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrileas a beige solid, mp 196-198° C.

MS (ES, positive ion mode): m/z calcd for C₃₃H₃₈ClN₇O₂S: 631.3, found:631.8, 633.7 (M+1).

EXAMPLE 344-(2,4-Dichloro-5-methoxyanilino)-6-methoxy-7-{3-[4-(1-pyrrolidinyl)-1-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrile

A mixture of 250 mg (0.536 mmol) of7-(3-chloropropoxy)-4-(2,4-dichloro-5-methoxyphenylamino)-6-methoxyquinoline-3-carbonitrile(Boschelli, Diane H.; Ye, Fei; Wang, Yanong D.; Dutia, Minu; Johnson,Steve L.; Wu, Biqi; Miller, Karen; Powell, Dennis W.; Yaczko, Deanna;Young, Mairead; Tischler, Mark; Arndt, Kim; Discafani, Carolyn; Etienne,Carlo, Gibbons, Jay; Grod, Janet; Lucas, Judy; Weber, Jennifer M.;Boschelli, Frank. J. Med. Chem. 2001, 44, 3965-3977) and 410 mg (2.68mmol) of 4-pyrrolidin-1-yl-piperidine are heated in 2 mL of ethyleneglycol dimethyl ether at 90° C. for 30 hours to yield the crude product.After adding water to the reaction mixture, the layers are separated.The organic layer is dried over sodium sulfate, which is removed byfiltration. Following removal of the solvent in vacuo, the residue isrecrystallized from ethyl acetate and ether to give 135 mg of4-(2,4-dichloro-5-methoxyanilino)-6-methoxy-7-{-[4-(1-pyrrolidinyl)-1-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrileas an off-white solid, mp 108-112° C.

MS (ES, positive ion mode): m/z calcd for C₃₀H₃₅Cl₂N₅O₃: 584.5, found:584.4; 586.3 (M+1).

EXAMPLE 357-(3-[1,4′-Bipiperidin]-1′-ylpropoxy)-4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-3-quinolinecarbonitrile

A mixture of 260 mg (0.56 mmol) of7-(3-chloropropoxy)-4-(2,4-dichloro-5-methoxyphenylamino)-6-methoxyquinoline-3-carbonitrile(Boschelli, Diane H.; Ye, Fei; Wang, Yanong D.; Dutia, Minu; Johnson,Steve L.; Wu, Biqi; Miller, Karen; Powell, Dennis W.; Yaczko, Deanna;Young, Mairead; Tischler, Mark; Arndt, Kim; Discafani, Carolyn; Etienne,Carlo; Gibbons, Jay; Grod, Janet; Lucas, Judy; Weber, Jennifer M.;Boschelli, Frank. J. Med. Chem. 2001, 44, 3965-3977) and 380 mg (2.24mmol) of [1,4′]bipiperidinyl are heated in 3 mL of ethylene glycoldimethyl ether at 85° C. for sixteen hours to yield the crude product.Following removal of the solvent in vacuo, the crude product is purifiedby silica gel chromatography (eluting with 7:3 methylenechloride/methanol) to give 35 mg of7-(3-[1,4′-bipiperidin]-1′-ylpropoxy)-4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-3-quinolinecarbonitrileas a white solid, mp 130-132° C.

MS (ES, positive ion mode): m/z calcd for C₃₁H₃₇Cl₂N₅O₃: 598.6, found:598.3; 600.3 (M+1).

EXAMPLE 364-{3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{2-[4-(1-pyrrolidinyl)-1-piperidinyl]ethoxy}-3-quinolinecarbonitrile

A mixture of 200 mg (0.40 mmol) of7-(2-chloroethoxy)-4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-3-quinolinecarbonitrileand 1.5 g (10 mmol) of 4-pyrrolidin-1-yl-piperidine are heated inN,N-dimethylformamide at 85° C. for 5 hours and at 40° C. for 60 hours.The reaction mixture is poured slowly into saturated aqueous sodiumbicarbonate to yield a solid which is subsequently filtered off. Thecrude solid is washed with water, then recrystallized from anhydrousethyl ether to give 203 mg of4-{3-chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{2-[4-(1-pyrrolidinyl)-1-piperidinyl]ethoxy}-3-quinolinecarbonitrileas an off-white solid, mp 127-130° C.

MS (ES, positive ion mode): m/z calcd for C₃₂H₃₆ClN₇O₂S: 618.2, found:617.8, 618.7 (M+1).

1-21. (canceled)
 22. A compound of Formula 1

wherein: Ar is a radical of the form:

wherein; A′ is a phenyl ring; wherein the phenyl ring is optionallymono- or di-substituted with a substituent selected from an alkyl of 1-6carbon atoms, an alkenyl of 2-6 carbon atoms, an alkynyl of 2-6 carbonatoms, azido, a hydroxyalkyl of 1-6 carbon atoms, a halogen, ahalomethyl, an alkoxymethyl of 2-7 carbon atoms, an alkanoyloxymethyl of2-7 carbon atoms, an alkoxy of 1-6 carbon atoms, an alkylthio of 1-6carbon atoms, hydroxy, a trifluoromethyl, a cyano, a nitro, a carboxy,an alkoxycarbonyl of 2-7 carbon atoms, an alkanoyl of 2-7 carbon atoms,a phenoxy, a phenyl, a thiophenoxy, a benzoyl, a benzyl, an amino, analkylamino of 1-6 carbon atoms, a dialkylamino of 2 to 12 carbon atoms,a phenylamino, a benzylamino, an alkanoylamino of 1-6 carbon atoms, analkenoylamino of 3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms,a carboxyalkyl of 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbonatoms, an aminoalkyl of 1-5 carbon atoms, a N-alkylaminoalkyl of 2-9carbon atoms, a N,N-dialkylaminoalkyl of 3-10 carbon atoms, anN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, a mercapto, a methylmercapto, an alkanoyloxy of 1-6 carbonatoms, a alkenoyloxy of 3-8 carbon atoms, an alkynoyloxy of 3-8 carbonatoms, a carbamoyl, a N-alkylcarbamoyl of 2-7 carbon atoms, aN,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; T issubstituted at a carbon of the phenyl ring with —O(CH₂)_(m)—, or—S(CH₂)_(m)—; L is alkyl or is an imidazole, pyridyl or a phenyl ringwherein the imidazole, pyridyl or phenyl ring is optionally substitutedat a carbon or nitrogen with one, two, or three substituentsindependently selected from an alkyl of 1-6 carbon atoms, an alkenyl of2-6 carbon atoms, an alkynyl of 2-6 carbon atoms, azido, a hydroxyalkylof 1-6 carbon atoms, a halogen, a halomethyl, an alkoxymethyl of 2-7carbon atoms, an alkanoyloxymethyl of 2-7 carbon atoms, an alkoxy of 1-6carbon atoms, an alkylthio of 1-6 carbon atoms, a hydroxy, atrifluoromethyl, a cyano, a nitro, a carboxy, an alkoxycarbonyl of 2-7carbon atoms, an alkanoyl of 2-7 carbon atoms, a phenoxy, a phenyl, athiophenoxy, a benzoyl, a benzyl, an amino, an alkylamino of 1-6 carbonatoms, a dialkylamino of 2 to 12 carbon atoms, a phenylamino, abenzylamino, an alkanoylamino of 1-6 carbon atoms, an alkenoylamino of3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms, a carboxyalkylof 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbon atoms, anaminoalkyl of 1-5 carbon atoms, an N-alkylaminoalkyl of 2-9 carbonatoms, an N,N-dialkylaminoalkyl of 3-10 carbon atoms, aN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, a mercapto, a methylmercapto, an alkanoyloxy of 1-6 carbonatoms, an alkenoyloxy of 3-8 carbon atoms, an alkynoyloxy of 3-8 carbonatoms, a carbamoyl, an N-alkylcarbamoyl of 2-7 carbon atoms, aN,N-dialkylcarbamoyl of 3-13 carbon atoms, a benzoylamino, a 5- or6-membered heteroaryl ring where the heteroaryl ring contains 1 to 3heteroatoms selected from N, O, and S and where the heteroaryl ring maybe optionally mono- or di-substituted with a substituent selected from ahalogen, an oxo, a thiocarbonyl, an alkyl of 1-6 carbon atoms, analkenyl of 2-6 carbon atoms, an alkynyl of 2-6 carbon atoms, azido, ahydroxyalkyl of 1-6 carbon atoms, a halomethyl, an alkoxymethyl of 2-7carbon atoms, an alkanoyloxymethyl of 2-7 carbon atoms, an alkoxy of 1-6carbon atoms, an alkylthio of 1-6 carbon atoms, a hydroxy, atrifluoromethyl, a cyano, a nitro, a carboxy, an alkoxycarbonyl of 2-7carbon atoms, an alkanoyl of 2-7 carbon atoms, a phenoxy, a phenyl, athiophenoxy, a benzoyl, a benzyl, an amino, an alkylamino of 1-6 carbonatoms, a dialkylamino of 2 to 12 carbon atoms, a phenylamino, abenzylamino, an alkanoylamino of 1-6 carbon atoms, an alkenoylamino of3-8 carbon atoms, an alkynoylamino of 3-8 carbon atoms, a carboxyalkylof 2-7 carbon atoms, a carboalkoxyalkyl of 3-8 carbon atoms, anaminoalkyl of 1-5 carbon atoms, an N-alkylaminoalkyl of 2-9 carbonatoms, a N,N-dialkylaminoalkyl of 3-10 carbon atoms, aN-alkylaminoalkoxy of 3-9 carbon atoms, a N,N-dialkylaminoalkoxy of 4-10carbon atoms, a mercapto, a methylmercapto, an alkanoyloxy of 1-6 carbonatoms, an alkenoyloxy of 3-8 carbon atoms, an alkynoyloxy of 3-8 carbonatoms, a carbamoyl, an N-alkylcarbamoyl of 2-7 carbon atoms, aN,N-dialkylcarbamoyl of 3-13 carbon atoms, and a benzoylamino; m is 0; nis 0; X is NH; R is alkyl of 1-6 carbon atoms; R₁ and R₄ are a hydrogen;R₂ is a hydrogen, —OH or —OR, where —OR is optionally substituted with—OH or —C₁-C₆alkoxy; R₃ is -0-(CH₂)₁₋₃-piperidine or—O—(CH₂)₁₋₃-piperazine optionally linked to a second ring selected frompiperidine, piperazine, pyrrolidine, morpholine, or imidazole, wheresaid second ring is optionally substituted with R₆, and where said linkis a bond or divalent straight or branched C₁-C₄alkyl; R₆ is hydrogen,an alkyl of 1-6 carbon atoms, an alkenyl of 2-6 carbon atoms, an alkynylof 2-6 carbon atoms, a cycloalkyl of 1-6 carbon atoms, an alkanoyl of2-7 carbon atoms, a carbamoylalkyl of 2-7 carbon atoms, a hydroxyalkylof 1-6 carbon atoms, a hydroxycycloalkyl of 3-6 carbon atoms, acarboxyalkyl of 2-7 carbon atoms, pyrrolidine, piperidine, or imidazoleoptionally substituted with methyl; or a crystalline form or apharmaceutically acceptable salt thereof.
 23. A compound of claim 22wherein: R₁ and R₄ are hydrogen; R₂ is hydrogen, an alkoxy of one tofour carbon atoms, a fluorine, or a nitro; R₃ is -0-(ddd)₁₋₃-piperidineor -0-(CH₂)₁₋₃-piperazine optionally substituted with pyrrolidine,piperidine, morpholine, imidazole optionally substituted with methyl; Xis NH; n is 0; Ar is a radical of the form:

wherein; A′ is a phenyl ring, optionally mono or di substituted with ahalogen; T is substituted at a carbon of the phenyl ring with—S(CH₂)_(m)— or —O(CH₂)_(m)—; m is 0; L is alkyl or is imidazole,optionally substituted on a nitrogen with a methyl or a phenyl ringoptionally substituted with one, two, or three substituentsindependently selected from an alkoxy of one carbon atom, a 5 or 6membered heteroaryl ring wherein the heteroaryl ring contains 1 or 2heteroatoms of N; or a crystalline form or a pharmaceutically acceptablesalt thereof.
 24. A compound of claim 22 wherein: R₁ and R₄ arehydrogen; R₂ is an alkoxy of one carbon atom; R₃ is piperidinesubstituted with pyrrolidine; X is NH; n is 0; Ar is a radical of theform:

wherein; A′ is a phenyl ring optionally mono or di substituted with achloride; T is substituted at a carbon of the phenyl ring with—S(CH₂)_(m)—, m is 0; L is alkyl or is imidazole, optionally substitutedon a nitrogen with a methyl; or a crystalline form or a pharmaceuticallyacceptable salt thereof.
 25. A compound selected from7-(2-[1,4′]Bipiperidinyl-1′-yl-ethoxy)-6-methoxy-4-(4-phenoxy-phenylamino)-quinoline-3-carbonitrile,4-{3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrile,4-(2,4-Dichloro-5-methoxyanilino)-6-methoxy-7-{3-[4-(1-pyrrolidinyl-1-7-{3-[4-(1-pyrrolidinyl)-1-piperidinyl]propoxy}-3-quinolinecarbonitrile,7-(3-[1,4′-Bipiperidin]-1′-ylpropoxy)-4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-3-quinolinecarbonitrile,and4-{3-Chloro-4-[(1-methyl-1H-imidazole-2-yl)sulfanyl]anilino}-6-methoxy-7-{2-[4(1-pyrrolidinyl)-1-piperidinyl]ethoxy}-3-quinolinecarbonitrile, or acrystalline form or a pharmaceutically acceptable salt thereof.
 26. Themethod of treating a melanoma, a pancreatic cancer, a colon cancer and alung cancer in a mammal comprising administering a compound of formula22.
 27. The method of claim 26 further comprising providing the mammalwith an effective amount of at least one anti-cancer agent.
 28. Themethod of claim 26 wherein the at least one anti-cancer agent isprovided prior to the at least one pharmaceutical composition,concurrently with the at least one pharmaceutical composition or afterthe at least one pharmaceutical composition.
 29. A means for preparing acompound of Formula I, of claim 22 comprising: a. reacting a compound ofFormula 2

with heated R₃—H, where H is bound to R₃ via a nitrogen atom; optionallywith a base; b. to yield the compound of Formula 1 of claim 1,

wherein: X′ is fluoro or chloro provided that when X′ is chloro, R₂ isnitro.